KR20220061967A - DcR3 variant - Google Patents

Abstract

The present invention has a ligand-binding activity (preferably neutralizing activity) of DcR3, and the amount of aggregates produced is reduced and/or improved when prepared using mammalian-derived cells as a host compared to wild-type DcR3. DcR3 variant showing in vivo kinetics, DNA encoding the DcR3 variant, a vector including the DNA, a transformant obtained by introducing the vector, a method for producing a variant using the transformant, and the individual An object of the present invention is to provide a preventive or therapeutic agent for autoimmune diseases, inflammatory diseases, or allergic diseases using the mutant as an active ingredient. provide a variant.

Description

DcR3 variant

The present invention relates to a DcR3 variant, which is a variant of wild-type DcR3. More specifically, the present invention has a ligand-binding activity (preferably neutralizing activity) of DcR3, and the production amount of aggregates is reduced compared to that of wild-type DcR3 when prepared using mammalian-derived cells as a host, And / or relates to a DcR3 variant showing improved in vivo kinetics.

Tumor necrosis factor (TNF) superfamily (TNFSF) and TNF receptor superfamily (TNFRSF) form structurally similar 18 ligands and 29 receptor families, respectively. Antibodies and Fc fusion proteins against many molecules included in this family have been developed and marketed, and have shown therapeutic effects in the treatment of various autoimmune diseases (Non-Patent Document 1).

Most of TNFRSF is expressed on the cell membrane and transmits signals downstream by ligand binding, but a few molecules are not involved in signal transduction, decoy (decoy) receptors (DcRs). Four decoy receptors have been identified: DcR1, DcR2, DcR3, and OPG (Osteoprotegerin).

OPG is a soluble decoy receptor for RANKL and TRAIL, and inhibits signal transduction by competing for binding with ligands of RANKL receptor and TRAIL receptor. On the other hand, DcR1 and DcR2 are decoy receptors that neutralize ligands by competitively inhibiting binding to signal transducing receptors, with respect to TRAIL and DcR3 for three molecules of LIGHT, TL1A and FasL (Non-Patent Document) 2).

DcR3 is a soluble molecule comprising 300 amino acid residues. On the N-terminal side, it has a signal peptide, followed by four cysteine-rich domains (CRDs) characteristic of TNFRSF (CRD1, CRD2, CRD3 and CRD4), and on the C-terminal side is heparan sulfate rich in basic amino acids containing a heparan sulfate binding motif. It has a bonding region (HBD). LIGHT, TL1A and FasL are all bound through CRD2 and CRD3 of DcR3 (Non-Patent Documents 3, 4, 5).

In addition to the function as a decoy receptor by ligand neutralization, DcR3 has a function as an immunomodulatory molecule based on HBD activity. For example, direct binding through HBD to glycosaminoglycans (GAG) including heparan sulfate on the cell membrane of monocytes, macrophages, or dendritic cells, Th2 induction by differentiation of dendritic cells, induction of M2 macrophages, It has been reported to induce various immune control and immunostimulating actions, such as enhancement of adhesion of monocytes, differentiation of osteoclasts, or reduction of expression of MHC class II molecules (Non-Patent Documents 6 and 12).

DcR3 ligand has been reported to be involved in autoimmune diseases, inflammatory diseases, allergies, cancer, infectious diseases, or other various inflammatory responses. For example, LIGHT, TL1A, and FasL are all included in the susceptibility locus in inflammatory bowel disease (IBD), and in particular, the existence of multiple genetic polymorphisms related to the pathology is known for TL1A. In addition, there are reports of improvement in the expression of DcR3 ligand in the blood or tissue of an IBD patient and improvement of the condition by inhibition of DcR3 ligand in a mouse enteritis model (Non-Patent Documents 6 to 9).

Although the expression of DcR3 in normal human tissues is very low, its expression is induced by infection or tissue disorders. In addition, in various autoimmune diseases or inflammatory diseases such as IBD, systemic erythematosus (SLE), atopic dermatitis (AD), or rheumatoid arthritis (RA), it can be seen that the blood level of DcR3 is elevated. . In addition, although a DcR3 homolog has not been identified in mice, in mouse pathological models such as a type I diabetes model, a multiple sclerosis model, or a nephritis model, human DcR3 transgenic mice improve the condition, and plasmid or recombinant The drug efficacy by administration of DcR3 has been confirmed, respectively (Non-Patent Documents 6 and 10).

Genentech Corporation cloned the human DcR3 gene and showed that a fusion of DcR3 and human IgG1 Fc region binds to soluble human FasL and inhibits human FasL-dependent apoptosis in vitro (Patent Document 1).

Eli Lilly Co., Ltd. obtained FLINT, a protease-resistant DcR3 mutant, by one amino acid mutation (R218Q) of wild-type DcR3, and reported that the in vivo kinetics was improved in mice and monkeys than in wild-type DcR3. However, when wild-type DcR3 and FLINT are administered to cynomolgus monkeys by intravenous injection of 0.5 mg/kg, the blood half-lives are very short, 9 hours and 12.3 hours, respectively (Patent Documents 2 and 3, Non-Patent Document 11).

Japanese Patent No. 4303883 US Patent US 6,835,814 B1 US Patent US 6,965,012 B1

Nature Reviews Drug Discovery, 2013, 12: p.147-168 Nature Reviews Cancer, 2002. 2: p.420-430 Structure, 2011, 19: p.162-171 Structure, 2014, 22: p.1252-1262 Structure, 2016, 24: p.2016-2023 Biochemical Pharmacology, 2011, 81: p.838-847 Immunology, 2009, 128: p.451-458 P.N.A.S., 2006, 103: p.8441-8446 Am. J. Physiol. Gastrointest. Liver Physiol., 2003, 285: p. G754-G760 Journal of Biomedical Science, 2017, 24: 39 Drug Metabolism and Disposition, 2003, 31: p.502-507 J. Immunol., 2006, 176: p.173-180

FLINT, which is a modified amino acid of wild-type DcR3, has remarkably poor in vivo kinetics, and frequent administration is required as a recombinant preparation with a ligand-neutralizing mechanism of action, which is undesirable as a pharmaceutical. Therefore, the usefulness of a DcR3 variant capable of securing an administration interval of a certain period by improving in vivo kinetics is expected as a pharmaceutical.

In addition, as far as is known so far, when expressed, isolated and purified using mammalian-derived cells as a host, the amount of aggregate production is reduced, and no functional DcR3 variants having neutralizing activity against DcR3 ligands have been found.

The present invention has a ligand-binding activity (preferably neutralizing activity) of DcR3, and the amount of aggregates produced is lowered compared to that of wild-type DcR3 when a DcR3 protein is prepared using a mammalian-derived cell as a host, and/or DcR3 variant showing improved in vivo dynamics, DNA encoding the DcR3 variant, a vector including the DNA, a transformant obtained by introducing the vector, a method for producing a variant using the transformant, and An object of the present invention is to provide a pharmaceutical composition comprising the modified body as an active ingredient, and a prophylactic or therapeutic agent for autoimmune diseases, inflammatory diseases, or allergies.

In order to solve the said subject, this invention provides the following invention.

[1] Wild-type decoy receptor (Decoy Receptor) 3 (hereinafter abbreviated as DcR3) as a variant DcR3 variant, which shows improved in vivo dynamics than the wild-type DcR3, the DcR3 variant.

[2] The DcR3 variant according to [1], having one or more N-glycosidic bond complex sugar chains.

[3] The DcR3 variant according to [1] or [2], which has neutralizing activity against at least one of LIGHT, TL1A and FasL.

[4] The DcR3 variant according to any one of [1] to [3], which has neutralizing activity against all of LIGHT, TL1A and FasL.

[5] The DcR3 variant according to any one of [1] to [3], which does not have neutralizing activity with respect to FasL, and has neutralizing activity with respect to any one or more of LIGHT and TL1A.

[6] DcR3 variant according to any one of [1] to [3] and [5], which does not have neutralizing activity with respect to FasL, and has neutralizing activity with respect to LIGHT and TL1A.

[7] In the cysteine-rich region of wild-type DcR3, at least a part of the cysteine-rich domain (hereinafter abbreviated as CRD) of wild-type DcR3 is substituted with at least a part of the cysteine-rich domain of a TNF receptor superfamily molecule other than DcR3 amino acid sequence A first chimeric-type cysteine-rich region comprising a, or a second chimeric-type cysteine comprising an amino acid sequence in which 1 to 30 amino acids are deleted, substituted, inserted or added in the amino acid sequence of the first chimeric-type cysteine-rich region Containing a rich region, DcR3 variant.

[8] The DcR3 variant according to [7], having one or more N-glycosidic bond complex sugar chains.

[9] The DcR3 variant according to [7] or [8], which has neutralizing activity against at least one of LIGHT, TL1A and FasL.

[10] The DcR3 variant according to any one of [7] to [9], which has neutralizing activity against all of LIGHT, TL1A and FasL.

[11] The DcR3 variant according to any one of [7] to [9], which does not have neutralizing activity with respect to FasL, and has neutralizing activity against any one or more of LIGHT and TL1A.

[12] The DcR3 variant according to any one of [7] to [9] and [11], which does not have neutralizing activity with respect to FasL, and has neutralizing activity against LIGHT and TL1A.

[13] The DcR3 variant according to any one of [7] to [12], wherein the TNF receptor superfamily molecule is OPG.

[14] At least a portion of the cysteine-rich domain of wild-type DcR3 is selected from all or part of CRD1, all or part of CRD2, all or part of CRD3, and all or part of CRD4,

At least a portion of the cysteine-rich domain of the TNF receptor superfamily molecule is selected from all or part of CRD1, all or part of CRD2, all or part of CRD3, and all or part of CRD4, [7] to [13] DcR3 variant described in any one.

[15] The first chimeric cysteine-rich region,

a substitution of a portion of CRD1 of the wild-type DcR3 in a portion corresponding to a portion of CRD1 of the wild-type DcR3 in CRD1 of the TNF receptor superfamily molecule;

substitution of all of CRD1 of said wild-type DcR3 in all of CRD1 of said TNF receptor superfamily molecule;

a substitution of a portion of CRD2 of said wild-type DcR3 in a portion corresponding to a portion of CRD2 of said wild-type DcR3 in CRD2 of said TNF receptor superfamily molecule;

substitution of all of CRD2 of said wild-type DcR3 in all of CRD2 of said TNF receptor superfamily molecule;

a substitution of a portion of CRD3 of said wild-type DcR3 in a portion corresponding to a portion of CRD3 of said wild-type DcR3 in CRD3 of said TNF receptor superfamily molecule;

substitution of all of CRD3 of said wild-type DcR3 in all of CRD3 of said TNF receptor superfamily molecule;

a substitution of a portion of CRD4 of said wild-type DcR3 in a portion corresponding to a portion of CRD4 of said wild-type DcR3 in CRD4 of said TNF receptor superfamily molecule, and

Substitution of all of CRD4 of said wild-type DcR3 in all of CRD4 of said TNF receptor superfamily molecule

Having one or more substitutions selected from, the DcR3 variant described in [14].

[16] The DcR3 variant according to [15], wherein in the first chimeric cysteine-rich region, part or all of CRD2 of the wild-type DcR3 is maintained.

[17] The DcR3 variant according to [15] or [16], wherein in the first chimeric cysteine-rich region, part or all of CRD3 of the wild-type DcR3 is maintained.

[18] The first chimeric type cysteine-rich region includes the following amino acid sequence (a), (b), (c) or (d), and the second chimeric type cysteine-rich region includes the following ( The DcR3 variant according to any one of [14] to [17], comprising the amino acid sequence of e).

(a) In the amino acid sequence of the cysteine-rich region of wild-type DcR3, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG

(b) in the amino acid sequence of the cysteine-rich region of wild-type DcR3, CRD4 of wild-type DcR3 is substituted with CRD4 of OPG

(c) an amino acid sequence in which CRD1 of wild-type DcR3 is substituted with CRD1 of OPG in the amino acid sequence of the cysteine-rich region of wild-type DcR3, and CRD4 of wild-type DcR3 is substituted with CRD4 of OPG

(d) in the amino acid sequence of (a), (b) or (c), the 103rd to 123rd position from the N-terminus is substituted with a corresponding part in the amino acid sequence of the cysteine-rich domain of OPG

(e) in the amino acid sequence of (a), (b), (c) or (d), 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence

[19] the amino acid sequence of (a) is an amino acid sequence comprising the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 26 or 50,

The amino acid sequence of (b) is an amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 28 or 52,

The amino acid sequence of (c) is an amino acid sequence comprising the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30 or 54,

The DcR3 variant according to [18], wherein the amino acid sequence of (d) is an amino acid sequence containing the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 32 or 56.

[20] The amino acid sequence of (e) is, from the N-terminus of the amino acid sequence of (a), (b), (c) or (d), Glu at position 57, Arg at position 58, and Arg at position 60 The DcR3 variant according to [18] or [19], wherein one or two or more amino acids selected from the group consisting of are substituted with other amino acids.

[21] The amino acid sequence of (e) is, from the N-terminus of the amino acid sequence of (a), (b), (c) or (d), Glu at position 57 and Arg at position 58 to another amino acid Having a substitution, the DcR3 variant according to any one of [18] to [20].

[22] The amino acid sequence of (e) above is Lys, Leu, Arg, Val, Ala of Glu at position 57 from the N-terminus of the amino acid sequence of (a), (b), (c) or (d) [18] having one or more substitutions selected from the group consisting of Phe, His, He or Met, Arg at position 58 with Asp, Glu, or Thr, and Arg at position 60 with Lys [18] ] to [21], the DcR3 variant according to any one of.

[23] The amino acid sequence of (e) above is Lys, Leu, Arg, Val, Ala of Glu at 57th position from the N-terminus of the amino acid sequence of (a), (b), (c) or (d) above; The DcR3 variant according to any one of [18] to [22], having a substitution with Phe, His, Ile or Met, and a substitution of Arg at the 58th position from the N-terminus with Asp, Glu or Thr.

[24] The DcR3 variant according to any one of [18] to [23], wherein the amino acid sequence of (e) has a substitution selected from the following (f) to (i).

(f) Substitution of Asn at positions 131 and 144 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid

(g) Substitution of Asn at positions 131, 144, and 157 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid

(h) Substitution of Thr at position 133 and Ser at position 146 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid

(i) Substitution of Thr at position 133, Ser at position 146, and Thr at position 159 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid

[25] The DcR3 variant according to any one of [18] to [24], wherein the amino acid sequence of (e) has a substitution selected from the following (f') to (i').

(f') Substitution of Asn at positions 131 and 144 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ser

(g') Substitution of Asn at positions 131, 144, and 157 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ser

(h') Substitution of Thr at position 133 and Ser at position 146 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ala

(i') Substitution of Thr at position 133, Ser at position 146, and Thr at position 159 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ala

[26] The amino acid sequence of (e) is SEQ ID NO: 58, 60, 62, 64, 66, 68, 70, 180, 182, 184, 186, 188, 270, 272, 274, 276, 278, 280, The DcR3 variant according to any one of [18] to [25], which is an amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus among the amino acid sequences described in 282, 284, or 286.

[27] A part of the heparan sulfate-binding region of the wild-type DcR3 wherein the DcR3 variant is bound to the C-terminal side of the first or second chimeric cysteine-rich region and the first or second chimeric cysteine-rich region; Among [7] to [26], it is a DcR3 variant including all or a DcR3 variant that contains the first or second chimeric cysteine-rich region and does not include the heparan sulfate binding region of the wild-type DcR3 DcR3 variant described in any one.

[28] The DcR3 variant,

(I) the amino acid sequence set forth in SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44 or 46, or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence, and

(II) SEQ ID NOs: 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 180, 182, 184, 186, 188, 270, 272, 274, 276, 278, 280, 282 , 284 or 286, or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence

The DcR3 variant according to [27], comprising any one amino acid sequence selected from.

[29] The DcR3 variant is a human IgG1, IgG2 or IgG4 antibody-derived Fc region, or in the amino acid sequence of the Fc region, one or several amino acids are deleted, substituted, inserted, or a mutation containing an amino acid sequence added DcR3 variants according to [7] to [28], including a type Fc region.

[30] The DcR3 variant according to [29], wherein the Fc region or the variant Fc region is linked to the C-terminal side of the first or second chimeric cysteine-rich region via another region or a linker. .

[31] The DcR3 variant according to [29] or [30], wherein the mutant Fc region has Ser at position 220 represented by the EU index in the amino acid sequence of the heavy chain of human IgG1.

[32] The mutant Fc region is, in the amino acid sequence of the human IgG1 heavy chain, a substitution of Ala for Leu at position 234, a substitution of Leu at position 235 with Ala, and a substitution of Gly at position 237 with Ala in the EU index Having a substitution, the DcR3 variant described in [31].

[33] The DcR3 variant according to [31] or [32], wherein the mutant Fc region has a substitution of Ala for Asn at position 434 indicated by the EU index in the amino acid sequence of the heavy chain of human IgG1.

[34] The mutant Fc region is, in the amino acid sequence of the human IgG1 heavy chain, Met at position 252, represented by the EU index, with Tyr, Ser at position 254, Thr substitution, and Thr at position 256 with Glu Having a substitution, the DcR3 variant described in [31].

[35] In the amino acid sequence of the human IgG4 heavy chain, the mutant Fc region is, in the amino acid sequence of the human IgG4 heavy chain, Ser at position 228 represented by the EU index by Pro, Leu at position 235 by Glu, and Arg at position 409 by Lys Having a substitution, the DcR3 variant according to [29] or [30].

[36] The DcR3 variant comprises a variant Fc region comprising the amino acid sequence set forth in SEQ ID NO: 72, 74, 156, 158, 160, 162, 164, 166, 311, 312 or 313, [29] or The DcR3 variant described in [30].

[37] The DcR3 variant is SEQ ID NO: 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 150, 168, 170, 172, 174, 176, 178, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 288, 290, 292, 294, 296, 298, 300, 302, 304, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336 or 337 amino acid sequence, or the corresponding The DcR3 variant according to any one of [29] to [36], comprising an amino acid sequence in which 1 to 30 amino acids are deleted, substituted, inserted or added in the amino acid sequence.

[38] A DcR3 variant composition comprising the variant DcR3 according to any one of [1] to [37].

[39] The composition according to [38], comprising a DcR3 variant having one or more N-glycosidic conjugated sugar chains and a DcR3 variant not having an N-glycosidic conjugated sugar chain.

[40] DNA encoding the DcR3 variant according to any one of [1] to [37].

[41] A genetic recombinant vector containing the DNA according to [40].

[42] A transformant obtained by introducing the recombinant vector according to [41] into a host cell.

[43] The transformant according to [42], wherein the host cell is a cell derived from a mammal.

[44] The transformant according to [43], wherein the mammal-derived cells are CHO cells.

[45] The transformant according to any one of [42] to [44] is cultured in a medium, and the DcR3 variant according to any one of [1] to [37] is produced and accumulated, and from the obtained culture medium, the DcR3 A method for producing a DcR3 variant or a DcR3 variant composition, characterized in that the variant is purified.

[46] A modified DcR3 or a modified DcR3 composition produced using the production method described in [45].

[47] A pharmaceutical composition comprising the modified DcR3 or modified DcR3 composition according to any one of [1] to [39] and [46] as an active ingredient.

[48] The pharmaceutical composition according to [47], which is a prophylactic or therapeutic agent for an autoimmune disease, an inflammatory disease, or an allergic disease.

[49] Autoimmune disease, inflammatory disease or allergic disease, comprising administering the pharmaceutical composition according to [47] or [48] to a patient in need of prevention or treatment of an autoimmune disease, inflammatory disease or allergic disease of prevention or treatment methods.

According to the present invention, DcR3 has a ligand-binding activity (preferably neutralizing activity), and when a DcR3 protein is prepared using a mammalian cell as a host, the amount of aggregates produced is reduced compared to that of wild-type DcR3, and / or a DcR3 variant showing improved in vivo dynamics, a DNA encoding the DcR3 variant, a vector including the DNA, a transformant obtained by introducing the vector, and a method for producing a variant using the transformant , and a pharmaceutical composition comprising the variant as an active ingredient, and a prophylactic or therapeutic agent for autoimmune diseases, inflammatory diseases, or allergies.

1A shows the results of SDS-PAGE of various wild-type DcR3 controls prepared from mammalian cells. In lanes 1 and 3, DcR3 FL-Fc, lanes 2 and 4, DcR3 FL-FLAG, and lanes 5 and 6, electrophoresis of S195-Fc (g1S) under non-reducing or reducing conditions, respectively. Fig. 1B shows that commercially available human DcR3-Fc (lane 7) prepared from HEK293 cells was electrophoresed under non-reducing conditions and detected by immunoblot with an anti-human IgG antibody. 1C shows commercially available DcR3 FL-Fc (lanes 8 and 9), S195-Fc (g1S) (lanes 10 and 11), and DcR3 FL-Fc (g1S) (lanes 14, 15, 18) prepared from insect cells. , 19) and R218Q-Fc(g1S) (lanes 12, 13, 16, 17) were electrophoresed under non-reducing or reducing conditions, respectively. M represents a molecular weight marker (Bio-Rad).
Fig. 2 shows the cysteine-rich domains as CRD1, CRD2, CRD3, and CRD4, respectively, in the alignment of the immature amino acid sequences of human DcR3 and human OPG.
3 schematically shows the domain structures of various wild-type DcR3 controls, various DcR3 variants, and human OPG produced. A: DcR3 FL-Fc, B: S195-Fc, C: chimeric A-Fc, D: 103-123OPG-Fc, E: N-type sugar chain 2 substituent-Fc, F: N-type sugar chain 3 substituent-Fc, G: Chimeric B-Fc, H: Chimeric C-Fc, I: OPG. The vertical lines in CRD4 of E and F indicate that there are 2 or 3 substitutions of the N-type sugar chain addition residue, respectively, and DD of I indicates the death domain.
4 shows the results of SDS-PAGE of various DcR3 variants prepared in mammalian cells. Lane 1 and 4 are chimeric A-Fc (IEGRMD g1S) prepared using Expi293 cells, lanes 2 and 5 are chimeric A-Fc (IEGRMD g1S) prepared using CHO-S cells, and lanes 3 and 6 are Expi293 cells. Chimeric B-Fc (g1S), lanes 7 and 8, was electrophoresed on chimeric C-Fc (IEGRMD g1S) prepared from Expi293 cells under non-reducing or reducing conditions. M represents a molecular weight marker (Bio-Rad).
Fig. 5 shows melting curves by the DSF method for R218Q-Fc, S195-Fc and chimeric A-Fc (IEGRMD g1S). The vertical axis indicates the fluorescence intensity RFU (10 ³ ), and the horizontal axis indicates temperature (°C).
6 shows the binding of various wild-type DcR3 controls and various DcR3 mutants to human primary cells and CHO cells. Each cell was reacted with each DcR3 variant at 10 µg/mL, then stained with a PE-labeled anti-human antibody at 0.1 µg/mL, and the fluorescence intensity of PE was measured by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE. The top shows HUVEC, the middle shows hepatocytes, and the bottom shows the staining results of CHO cells.
Fig. 7 shows the transition of blood concentration in BALB/c mice when S195-Fc and chimeric A-Fc (IEGRMD g1S) were administered at 10 mg/kg iv. The vertical axis represents the blood concentration (ng/mL), and the horizontal axis represents the elapsed time (hr) after administration.
8A shows the binding of various wild-type DcR3 controls and various DcR3 variants to RANKL (OPG ligand) and FIG. 8B to TRAIL (OPG ligand), respectively. After capturing various wild-type DcR3 controls and various DcR3 variants on a plate immobilized with an anti-human antibody, RANKL or TRAIL diluted to each concentration was added to evaluate binding. A biotinylated anti-RANKL antibody or a biotinylated anti-TRAIL antibody and streptavidin-HRP were used for detection. The horizontal axis represents the concentration (pg/mL) of RANKL or TRAIL, and the vertical axis represents the absorbance (value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm).
9 shows the neutralizing activity of various wild-type DcR3 controls and various DcR3 variants for LIGHT. LIGHT 100ng/mL, and various wild-type DcR3 controls and various DcR3 variants 0.1, 1, or 10㎍/mL is added, IL-8 production from HT-29 cells is shown. The vertical axis indicates the IL-8 concentration (pg/mL), and the horizontal axis indicates various DcR3 variants added as an inhibitor.
10 shows the neutralizing activity of various wild-type DcR3 controls and various DcR3 variants for TL1A. IFN-γ production from human T cells is shown when TL1A is added at 100 ng/mL, and various wild-type DcR3 controls and various DcR3 variants are added at 0.1, 1, or 10 μg/mL. The vertical axis indicates the IFN-γ concentration (pg/mL), and the horizontal axis indicates various DcR3 variants added as an inhibitor.
11 shows the neutralizing activity of various wild-type DcR3 and various DcR3 mutant controls for FasL. The amount of ATP produced from living Jurkat cells when FasL was added to 100 ng/mL and 0.01, 0.1, or 1 μg/mL of various wild-type DcR3 controls and various DcR3 variants was expressed as RLU. The vertical axis indicates cell survival (RLU×10 ⁶ ) using ATP-dependent chemiluminescence as an index, and the horizontal axis indicates various DcR3 variants added as an inhibitor.
Figure 12a shows the binding of the chimeric A-Fc (g4PEK) and the FasL binding lowering variant (g4PEK) to each DcR3 ligand. A sensor when a chimeric A-Fc (g4PEK) or each FasL binding-reducing variant is captured on a sensor chip immobilized with an anti-human antibody, and a DcR3 ligand (human FasL, human LIGHT, human TL1A) is flowed as an analyte. represents grams. The vertical axis represents the binding amount (RU), and the horizontal axis represents time (Sec).
Figure 12b shows the binding of the FasL binding lowering variant (g4PEK) to each DcR3 ligand. A sensorgram obtained when each FasL binding-reducing variant was captured on a sensor chip immobilized with an anti-human antibody and DcR3 ligand (human FasL, human LIGHT, human TL1A) was flowed as an analyte is shown. The vertical axis represents the binding amount (RU), and the horizontal axis represents time (Sec).
Figure 12c shows the binding of the FasL binding lowering variant (g4PEK) to each DcR3 ligand. A sensorgram obtained when each FasL binding-reducing variant was captured on a sensor chip immobilized with an anti-human antibody and DcR3 ligand (human FasL, human LIGHT, human TL1A) was flowed as an analyte is shown. The vertical axis represents the binding amount (RU), and the horizontal axis represents time (Sec).
Fig. 13 shows the results of SDS-PAGE of the wild-type DcR3 control prepared with various mammalian cells. SDS-PAGE was performed by electrophoresing commercially available human DcR3-Fc prepared from various mammalian cells under reducing or non-reducing conditions. As mammalian cells, HEK293 cells (Abcam) were used in lanes 1 and 4, CHO cells (AdipoGen) were used in lanes 2 and 5, and HEK293 cells (Enzo) were used in lanes 3 and 6. M represents a molecular weight marker (Bio-Rad).
Figure 14a shows the ratio (%) of the contents of monomers, aggregates and decomposition products calculated from the peak area of SEC-HPLC or SEC-UPLC of the protein A purified FasL binding lowering variant. Figure 14a shows the results of all the produced FasL binding lowering variants (g4PEK).
Figure 14b shows the ratio (%) of the content of monomers, aggregates and degradation products calculated from the peak area of SEC-HPLC or SEC-UPLC of the protein A purified FasL binding lowering variant. Figure 14b shows the results of various mutant Fc fusions of the selected FasL binding lowering variants.
Fig. 14C shows the ratio (%) of the contents of monomers, aggregates and degradation products calculated from the peak areas of SEC-HPLC or SEC-UPLC of various chimeric A-mutant Fc fusions purified by Protein A.
Figure 15a shows the results of measuring the binding activity of various DcR3 variants to each DcR3 ligand by BIAcore. Various kinetic constants (k _a , k _d , K _D ). 15A shows the results of various chimeric A-Fc having different Fc sequences.
Figure 15b shows the results of measuring the binding activity of various DcR3 variants to each DcR3 ligand by BIAcore. Various kinetic constants (k _a , k _d , K _D ). Figure 15b shows the results of the FasL binding lowering variant.
Figure 16a shows the results of comparing the kinetic constants (K _D values) for each DcR3 ligand calculated by BIAcore when selecting variants with reduced FasL binding to the chimeric A-Fc (g4PEK). Fig. 16A shows the results of one amino acid substitution.
Figure 16b shows the result of comparing the kinetic constants (K _D values) for each DcR3 ligand calculated by BIAcore when selecting variants with reduced FasL binding to the chimeric A-Fc (g4PEK). Fig. 16B shows the results of two amino acid substitutions.
Figure 17a shows the results of evaluation of neutralizing activity of various DcR3 variants for soluble LIGHT. Fig. 17a shows inhibitory activity on LIGHT-dependent-CXCL10 production from IFN-γ-stimulated intestinal myofibroblasts by various chimeric A-Fc having different Fc sequences. The vertical axis indicates the concentration of CXCL10 (ng/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 17b shows the results of evaluation of the neutralization activity of various DcR3 variants for soluble LIGHT. Fig. 17B shows inhibitory activity on LIGHT-dependent CXCL10 production from IFN-γ-stimulated intestinal myofibroblasts by chimeric A-Fc having a different mutated Fc sequence. The vertical axis indicates the concentration of CXCL10 (ng/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 17c shows the results of evaluation of neutralization activity of various DcR3 variants for soluble LIGHT. Figure 17c shows the inhibitory activity on LIGHT-dependent IL-8 production from HT-29 cells by the 1-amino acid substitution FasL binding lowering variant. The vertical axis indicates the IL-8 concentration (ng/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 17d shows the results of evaluation of neutralizing activity of various DcR3 variants for soluble LIGHT. Figure 17d shows the inhibitory activity on LIGHT-dependent CXCL10 production from IFN-γ-stimulated intestinal myofibroblasts by the 2-amino acid substitution FasL binding lowering variant. The vertical axis indicates the concentration of CXCL10 (ng/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 18a shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble TL1A. Fig. 18A shows inhibitory activity against TL1A-dependent IFN-γ production from IL-12 and IL-18-stimulated human T cells by various chimeric A-Fc having different Fc sequences. The vertical axis indicates the concentration of IFN-γ (pg/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 18b shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble TL1A. Fig. 18B shows the inhibitory activity on TL1A-dependent IFN-γ production from IL-12 and IL-18-stimulated human T cells by chimeric A-Fc having different mutated Fc sequences. The vertical axis indicates the concentration of IFN-γ (pg/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 18c shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble TL1A. Fig. 18c shows the inhibitory activity on TL1A-dependent IFN-γ production from IL-12 and IL-18-stimulated human T cells by the 1-amino acid substitution FasL binding-lowering variant. The vertical axis indicates the concentration of IFN-γ (pg/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 18d shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble TL1A. Figure 18d shows the inhibitory activity on TL1A-dependent IFN-γ production from IL-12 and IL-18-stimulated human T cells by the 2-amino acid substitution FasL binding lowering variant. The vertical axis indicates the concentration of IFN-γ (pg/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 19a shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble FasL. Fig. 19A shows the inhibitory activity against apoptosis of A3 cells by various chimeric A-Fc having different Fc sequences. The vertical axis indicates cell survival (RLU×10 ⁶ ) using ATP-dependent chemiluminescence as an index, and the horizontal axis indicates the concentration of various DcR3 variants added (ng/mL).
Figure 19b shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble FasL. 19B shows the inhibitory activity against cell death of Jurkat cells by the chimeric A-Fc having another mutated Fc sequence. The vertical axis indicates cell survival (RLU×10 ⁶ ) using ATP production as an index, and the horizontal axis indicates the concentration of various DcR3 variants added (ng/mL).
Figure 19c shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble FasL. Figure 19c shows the inhibitory activity against apoptosis of Jurkat cells by the 1-amino acid substitution FasL binding lowering variant. The vertical axis indicates cell survival (RLU×10 ⁶ ) using ATP production as an index, and the horizontal axis indicates the concentration of various DcR3 variants added (ng/mL).
Figure 19d shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble FasL. Figure 19d shows the inhibitory activity on cell death of Jurkat cells by the 2-amino acid substitution FasL binding lowering variant. The vertical axis indicates cell survival (RLU×10 ⁶ ) using ATP production as an index, and the horizontal axis indicates the concentration of various DcR3 variants added (ng/mL).
Fig. 20a shows the results of evaluation of binding activity of S195-Fc and various chimeric A-Fc having different Fc sequences to the membranous LIGHT forced expression line. Each cell was reacted with various DcR3 variants, stained with a PE-labeled anti-human antibody, and the fluorescence intensity of PE was measured by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Fig. 20B shows the results of evaluation of binding activity of S195-Fc and various chimeric A-Fc having different Fc sequences with respect to membrane-type TL1A and membrane-type FasL forced expression lines. Each cell was reacted with various DcR3 variants, stained with a PE-labeled anti-human antibody, and the fluorescence intensity of PE was measured by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Figure 21a shows the results of evaluation of binding activity of S195-Fc, various chimeric A-Fc, and FasL binding lowering variants for membranous LIGHT forced expression strains. Each cell was reacted with various DcR3 variants, stained with a PE-labeled anti-human antibody, and the fluorescence intensity of PE was measured by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Fig. 21b shows the results of evaluation of binding activity of S195-Fc, various chimeric A-Fc, and FasL binding lowering variants to the membranous TL1A forced expression strain. Each cell was reacted with various DcR3 variants, stained with a PE-labeled anti-human antibody, and the fluorescence intensity of PE was measured by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Figure 21c shows the results of evaluation of the binding activity of the chimeric A-Fc and FasL binding lowering variant (g4PEK) to the membranous FasL forced expression strain. Each cell was reacted with various DcR3 variants, stained with a PE-labeled anti-human antibody, and the fluorescence intensity of PE was measured by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Fig. 22 shows the results of evaluation of the binding activity of the chimeric A-Fc to the membrane-type LIGHT of the primary cells. Fig. 22A shows the results of measuring the expression of membranous LIGHT on activated human T cells with a PE-labeled anti-LIGHT antibody. Fig. 22B shows the geometric mean (Geo.Mean) calculated by measuring the binding of Alexa Fluor 488-labeled chimeric A-Fc to membrane-type LIGHT on activated human T cells by flow cytometry. In all cases, the vertical axis represents the geometric mean (Geo.Mean) of fluorescence intensity.
Fig. 23 shows the results of evaluation of the binding activity of the chimeric A-Fc to the membranous TL1A of the primary cells. Binding of Alexa Fluor 647-labeled chimeric A-Fc to membranous TL1A on HUVEC cells in the presence or absence of a competitive protein was measured by flow cytometry, respectively, and the calculated geometric mean (Geo.Mean) is shown. .
Fig. 24 shows the results of measuring the binding activity of the chimeric A-Fc to FasL derived from primary cells by sandwich ELISA for soluble FasL in the culture supernatant of AICD-induced human T cells. Fig. 24A shows chimeric A-Fc, and Fig. 24B shows the absorbance value at 450 nm when the culture supernatant is added to a plate in which Fas-Fc has been captured, and detected with an anti-FasL antibody.
Fig. 25 shows the results of elution time (minutes) in hydrophobic interaction chromatography (HIC) of various chimeric A-Fc and FasL binding lowering variants having different Fc sequences.
26 shows the results of Tm values (°C) calculated by Differential Scanning Fluorimetry (DSF) method of various chimeric A-Fc and FasL binding lowering variants having different Fc sequences.
Fig. 27 shows the blood half-life (h) and time to infinity of chimeric A-Fc (Eg1S) and FasL binding lowering variants having different Fc sequences in BALB/c mice after 10 mg/kg iv administration. The value of AUC0-∞ (μg*h·mL) of the area under the blood concentration-time curve is shown.
Fig. 28A shows the index of gross pathology score in a drug efficacy test of chimeric A-Fc using a mouse acute heterogeneous GVHD model.
28B shows the condition score for each individual in each group and the mean of each group. The ordinate represents the condition score, and the abscissa represents each treatment group.
Fig. 29 shows the number of cells per spleen for each of human CD45 positive, human CD3 and CD4 positive, human CD3 and CD8 positive cells in the drug efficacy test of chimeric A-Fc using a mouse acute heterogeneous GVHD model. It is expressed as the average value for each individual and each group. Group 1 represents no human cell transfection, group 2 represents a DNP antibody administration group with human cell transfection, and group 3 represents a chimeric A-Fc administration group with human cell translocation, respectively. The vertical axis indicates the number of cells positive for each surface marker, and the horizontal axis indicates each treatment group.
Figure 30a shows the results of measuring the binding activity of various DcR3 variants to human DcR3 ligand by BIAcore. Various kinetic constants (k _a , k _d , K _D ) are shown when a trimer of human FasL, human LIGHT, or human TL1A is used as a human DcR3 ligand.
Figure 30b shows the results of measuring the binding activity of various DcR3 variants to the cynomolgus monkey DcR3 ligand by BIAcore. Various kinetic constants (k _a , k _d , K _D ) are shown when a trimer of cynomolgus monkey FasL, cynomolgus monkey LIGHT, or cynomolgus monkey TL1A is used as the cynomolgus DcR3 ligand.
Figure 31 shows the results of evaluation of the neutralization activity of various DcR3 variants for soluble LIGHT. Figure 31 shows the inhibitory activity of LIGHT-dependent CXCL10 production of chimeric A-Fc and FasL binding-reducing variants having various mutated Fc. The vertical axis indicates the concentration of CXCL10 (ng/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 32 shows the results of evaluation of the neutralizing activity of various DcR3 variants for soluble TL1A. Figure 32 shows the inhibitory activity of the chimeric A-Fc and FasL binding lowering variants having various mutated Fc on TL1A-dependent IFN-γ production of human T cells. The vertical axis indicates the concentration of IFN-γ (pg/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
33 shows the results of evaluation of the neutralization activity of various DcR3 variants for soluble FasL. Figure 33 shows the inhibitory activity against the cell death of Jurkat cells by the chimeric A-Fc and FasL binding lowering variants having various mutated Fc. The vertical axis indicates cell survival (RLU×10 ⁶ ) using ATP-dependent chemiluminescence as an index, and the horizontal axis indicates the concentration of various DcR3 variants added (ng/mL).
Figure 34a shows the results of evaluating the binding activity of various DcR3 variants to the membranous LIGHT forced expression line by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Figure 34b shows the results of evaluation of the binding activity of various DcR3 variants to the membranous TL1A forced expression strain by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
Figure 34c shows the results of evaluation of the binding activity of various DcR3 variants to the membrane-type FasL forced expression strain by flow cytometry. The vertical axis of the graph represents the geometric mean (Geo.Mean) of PE.
35 shows the results of evaluation of the neutralization activity of various DcR3 variants for membranous LIGHT. Figure 35 shows the inhibitory activity of the chimeric A-Fc and FasL binding lowering variants having various mutated Fc on the membranous LIGHT-dependent CXCL10 production of intestinal myofibroblasts. The vertical axis indicates the concentration of CXCL10 (ng/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 36 shows the results of evaluation of the neutralizing activity of various DcR3 variants for membranous TL1A. Fig. 36 shows the inhibitory activity on membrane-type TL1A-dependent IFN-γ production of human CD4-positive T cells by the chimeric A-Fc and FasL binding-lowering variants having various mutated Fc. The vertical axis indicates the concentration of IFN-γ (pg/mL), and the horizontal axis indicates the concentration of the added DcR3 variant (ng/mL).
Figure 37 shows the results of evaluation of the neutralization activity of various DcR3 variants with respect to membrane FasL. Figure 37 shows the inhibitory activity against membrane-type FasL-dependent cell death of Jurkat cells by the chimeric A-Fc having various mutated Fc and the FasL binding lowering variant. The vertical axis of the graph indicates the percentage (%) of the dead cell fraction positive for Annexin V, and the horizontal axis indicates the concentration (ng/mL) of various DcR3 variants added.
Figure 38a shows the results of measuring the binding activity of various DcR3 variants to human LIGHT recombinant by sandwich ELISA. The ordinate represents the value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm, and the abscissa represents the concentration (ng/mL) of the added recombinant human LIGHT.
Figure 38b shows the results of measuring the binding activity of various DcR3 variants to LIGHT derived from human primary cells by sandwich ELISA for soluble LIGHT in culture supernatants of human T cells. The white bar shows the culture supernatant when human T cells are cultured without stimulation, and the black bar shows the results of using the culture supernatant of human T cells stimulated with anti-CD3 antibody and anti-CD28 antibody. The ordinate indicates a value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm.
Figure 38c shows the results of measuring the binding activity of various DcR3 variants to the recombinant human TL1A by sandwich ELISA. The vertical axis represents the value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm, and the horizontal axis represents the concentration (ng/mL) of the recombinant TL1A.
Fig. 38D shows the results of measuring the binding activity of various DcR3 variants to TL1A derived from human primary cells by sandwich ELISA for soluble TL1A in the culture supernatant of human PBMCs. The white bar shows the culture supernatant when human PBMCs were cultured without stimulation, and the black bar shows the results of using the culture supernatant of human PBMCs stimulated with immune complexes. The ordinate indicates a value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm.
Figure 38e shows the results of measuring the binding activity of various DcR3 variants to the recombinant human FasL by sandwich ELISA. The vertical axis represents the value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm, and the horizontal axis represents the human FasL concentration (ng/mL).
Fig. 38F shows the results of measuring the binding activity of various DcR3 variants to FasL derived from human primary cells by sandwich ELISA for soluble FasL in the culture supernatant of AICD-induced human T cells. The white bar shows the culture supernatant of non-AICD-induced human T cells, and the black bar shows the result of using the culture supernatant of AICD-induced human T cells. The vertical axis represents a value obtained by subtracting the absorbance at 570 nm from the absorbance at 450 nm.
Figure 39 shows the blood half-life of the disappearance phase (h) and the blood concentration-time curve to infinity after 10 mg/kg iv administration of various DcR3 variants to BALB/c mice, AUC0-∞ (μg*h·mL) represents the value of
40 shows the results of detection of various DcR3 variants transiently expressed in mammalian cells and not Fc-fused by immunoblot. Lanes 1 and 5 are 30-fold concentrates of the culture supernatant expressing S195-His6, lanes 2 and 6 are 6-fold dilutions of the culture supernatants expressing chimeric A-His6, and lanes 3 and 7 are culture supernatants expressing E57K-His6 of 6-fold dilutions, lanes 4 and 8, 6-fold dilutions of the culture supernatant expressing 45-18-His6 were electrophoresed under non-reducing or reducing conditions, respectively, and immunoblot with anti-6-His tag antibody. detected.

EMBODIMENT OF THE INVENTION Hereinafter, the suitable form for implementing this invention is demonstrated. In addition, embodiment described below shows an example of typical embodiment of this invention, and the scope of this invention is not interpreted narrowly by this.

Two or more of the forms described below may be combined, and such combinations are also included in the present invention.

The present invention relates to a DcR3 variant, which is a variant of wild-type DcR3. Specifically, the present invention relates to a DcR3 variant having DcR3 ligand binding activity (or neutralizing activity), and exhibiting improved in vivo kinetics compared to wild-type DcR3 and/or lowered aggregation than wild-type DcR3 when produced in mammalian cells, And in the cysteine-rich domain of wild-type DcR3, it relates to a DcR3 variant comprising a cysteine-rich domain in which a mutation is introduced in one or two or more amino acids.

1. Wild-type DcR3

DcR3 is also commonly referred to as decoy receptor 3, DCR3, TNFRSF6B (Tumor necrosis factor receptor superfamily member 6B), TR6 or M68. DcR3, which belongs to the TNF receptor superfamily, is a soluble decoy receptor that does not have a transmembrane domain, and by binding to three ligands of LIGHT, TL1A or FasL, competitively inhibits the binding of each ligand and the receptor, , to neutralize the ligand. In addition, as a function other than neutralization of the ligand, it directly binds to glycosaminoglycans (GAG) including heparan sulfate on the cell membrane of monocytes, macrophages, or dendritic cells through the heparan sulfate binding region (HBD), It has been reported to cause various immune control and immunostimulating actions [Biochemical. Pharmacology, 2011, 81: p. 838-847, J. Immunol., 2006, 176: p. 173-180].

DcR3 existing in nature has CRD1, CRD2, CRD3, CRD4 and HBD in order from the N-terminal side. DcR3 existing in nature further has a region present between CRD1 and CRD2, a region present between CRD2 and CRD3, a region present between CRD3 and CRD4, and a region present between CRD4 and HBD. The naturally occurring cysteine-rich region of DcR3 is a region from the N-terminus of CRD1 to the C-terminus of CRD4, including CRD1, CRD2, CRD3 and CRD4, and a region existing between CRD1 and CRD2, CRD2 and CRD3 a region between, and a region between CRD3 and CRD4. "Wild-type DcR3" is a molecule containing a cysteine-rich region and HBD, and is a molecule in which the cysteine-rich region and HBD are wild-type (that is, the cysteine-rich region and HBD are identical to the cysteine-rich region and HBD of naturally occurring DcR3). means Therefore, in addition to naturally occurring DcR3, "wild-type DcR3" also includes naturally occurring DcR3 variants such as genetic polymorphisms and isoforms, as long as they include the cysteine-rich region and HBD of naturally occurring DcR3. In addition, immature type DcR3 and mature type DcR3 are also included in "wild type DcR3". "Immature DcR3" means DcR3 having a signal peptide, and "mature DcR3" means DcR3 in which the signal peptide was cleaved. The signal peptide may be any of a sequence derived from naturally occurring DcR3, an artificial sequence, a sequence derived from an expression vector, and a sequence derived from another protein suitable for a host cell expressing naturally occurring DcR3. In addition, when the cleavage site differs depending on the signal peptide to be used, the sequence in which the N-terminal amino acid of the mature type differs is also included in "wild type DcR3". The CRD of naturally occurring DcR3 and the CRD of wild-type DcR3 are also referred to as “wild-type CRD”, and the naturally occurring cysteine-rich region and wild-type DcR3 cysteine-rich region are also referred to as “wild-type cysteine-rich region”.

Although the origin is not limited as for the wild-type DcR3 in this invention, DcR3 derived from various eukaryotes is mentioned. For example, DcR3 derived from amphibians such as frogs, birds such as chickens, or mammals, such as primates including humans, or oxen such as pigs and cattle. When the DcR3 variant of the present invention is used in humans, it is preferable to use human-derived DcR3 as the wild-type DcR3.

The cDNA sequence of human-derived DcR3 is shown in SEQ ID NO: 1, and the corresponding mRNA sequence is registered with GenBank (NCBI, USA) as accession number: NM_003823.3. The amino acid sequence of human-derived DcR3 is shown in SEQ ID NO: 2 and is registered with GenBank (NCBI, USA) as accession number: NP_003814.1. The immature human DcR3 has a signal peptide at the N terminus, followed by four CRDs (CRD1, CRD2, CRD3, CRD4) characteristic of the TNF receptor superfamily, and an HBD rich in basic amino acids at the C terminus. The mature human DcR3 is one in which the signal peptide was cleaved out of the immature form, and the amino acid sequence of the mature human DcR3 is, for example, shown in SEQ ID NO: 4 and is a DNA encoding the amino acid sequence of the mature human DcR3. The base sequence is shown, for example, in SEQ ID NO:3. In the present invention, in the amino acid sequence of human DcR3 (SEQ ID NO: 2), the region 30 to 70 from the N-terminus is CRD1 (SEQ ID NO: 6), the region 73 to 113 is CRD2 (SEQ ID NO: 8), 115 to The 150th region is defined as CRD3 (SEQ ID NO: 10), the 153rd to 193rd region is defined as CRD4 (SEQ ID NO: 12), and the 196th to 300th region is defined as HBD (SEQ ID NO: 48) (Fig. 2). The nucleotide sequences of DNA encoding the amino acid sequences of CRD1, CRD2, CRD3, CRD4 and HBD of human DcR3 are shown, for example, in SEQ ID NOs: 5, 7, 9, 11 and 47, respectively. In addition, the amino acid sequence of the CRD has a plurality of definitions other than those described above, but any definition of the amino acid sequence of the CRD can be used for the DcR3 variant of the present invention using known information [UniProt O95407, GenBank NP_003814.1, Structure, 2011, 19: p.162-171].

Examples of the DcR3 ligand include LIGHT, TL1A and FasL, all belonging to TNFSF as described above.

LIGHT (lymphotoxin-like, exhibits inducible expression, and competes with herpes simplex virus (HSV) glycoprotein D (gD) for HVEM, a receptor expressed by T lymphocytes, which exhibits inducible expression, and competes with herpes simplex The virus (HSV) glycoprotein D (gD) for HVEM, a receptor expressed by Tlymphocytes)) is commonly referred to as TNFSF14 (tumor necrosis factor superfamily member 14), LTg, HVEM-L, or CD258. The mRNA sequence of human LIGHT and its corresponding cDNA sequence are registered with GenBank (NCBI, USA) as accession number: NM_003807.4 and amino acid sequence as accession number: NP_003798.2. Soluble LIGHT is produced by expressing on the cell membrane as membrane-type LIGHT and then shedding the extracellular region with a protease. The cleavage site in membranous LIGHT is between the 82nd and 83rd amino acids of NP_003798.2. Both the soluble type and the membrane type are functional.

TL1A (tumor necrosis factor (TNF)-like cytokine 1A) is also commonly referred to as TNFSF15 (TNF superfamily member 15), TL1, VEGI or VEGI-251. The mRNA sequence of human TL1A and its corresponding cDNA sequence are registered with GenBank (NCBI, USA) as accession number: NM_005118.3 and amino acid sequence as accession number: NP_005109.2. Soluble TL1A is produced by expressing on a cell membrane as membrane-type TL1A and then shedding the extracellular region with a protease. The cleavage site in membrane-type TL1A is between the 71st and 72nd amino acids of NP_005109.2, and both soluble and membrane-type are functional.

FasL (Fas ligand) is also commonly referred to as FASLG, TNFSF6 (tumor necrosis factor superfamily member 6), CD178 or APT1LG1. The mRNA sequence of human FasL and its corresponding cDNA sequence are registered with GenBank (NCBI, USA) as accession number: NM_000639.2 and amino acid sequence as accession number: NP_000630.1. Soluble FasL is produced by expressing on a cell membrane as membrane FasL and then shedding the extracellular region with a protease. The cleavage site in membrane FasL is between positions 81 and 82 or between positions 129 and 130 of NP_000630.1. In vivo, membrane-type FasL has been mainly reported as a functional ligand.

In addition, polymorphisms or isoforms of genes are often identified in genes encoding eukaryotic proteins. In the gene used in the present invention, a gene in which a mutation in a nucleotide sequence or an amino acid sequence is caused by such a polymorphism is also included in the gene encoding LIGHT, TL1A or FasL in the present invention.

2. DcR3 variant

The DcR3 variant of the present invention includes a chimeric cysteine-rich region.

2-1. Chimeric cysteine rich region

The chimeric cysteine-rich region of the present invention includes an amino acid sequence in which one or two or more amino acids are mutated in the amino acid sequence of the cysteine-rich region of wild-type DcR3. In addition, "a mutation is introduced into a certain amino acid sequence/base sequence" means that one or two or more amino acids/bases are substituted, deleted, inserted, or added in the sequence. "Substitution, deletion, insertion or addition" also includes a combination of two or more mutations selected from substitution, deletion, insertion and addition. The mutation is introduced into one or two or more CRDs selected from at least CRD1, CRD2, CRD3 and CRD4 among the cysteine-rich regions of wild-type DcR3. Of the cysteine-rich regions of wild-type DcR3, mutations may or may not be introduced into the region between CRD1 and CRD2, the region between CRD2 and CRD3, and the region between CRD3 and CRD4. When a mutation is introduced into one or two or more regions selected from the region between CRD1 and CRD2, the region between CRD2 and CRD3, and the region between CRD3 and CRD4, each region after the mutation is introduced The number of amino acids is usually 1 to 10, preferably 1 to 7, still more preferably 1 to 5, still more preferably 1 to 3, still more preferably 1 to 2. The amino acid sequence of each region after the mutation is introduced is not particularly limited. Mutations introduced into the cysteine-rich region of wild-type DcR3 include both naturally occurring mutations and artificial mutations. The chimeric cysteine-rich region in the present invention includes, for example, an amino acid sequence in which one or two or more amino acids are substituted, deleted, inserted or added in the amino acid sequence of the cysteine-rich region of wild-type DcR3. region, and examples of such a chimeric type cysteine-rich region include, for example, first and second chimeric type cysteine-rich regions described later.

2-1-1. first chimeric cysteine rich region

The first chimeric cysteine-rich region includes an amino acid sequence in which at least a part of CRD of wild-type DcR3 is substituted with another peptide or protein in the amino acid sequence of the cysteine-rich region of wild-type DcR3. That is, the first chimeric type cysteine-rich region includes an amino acid sequence derived from the cysteine-rich region of wild-type DcR3 and an amino acid sequence derived from another peptide or protein.

Among the cysteine-rich regions of wild-type DcR3, the portion substituted with another peptide or protein is preferably at least a portion of at least one CRD selected from CRD1, CRD2, CRD3 and CRD4. Accordingly, in the cysteine-rich region of wild-type DcR3, a portion substituted with another peptide or protein may be selected from all or part of CRD1, all or part of CRD2, all or part of CRD3, and all or part of CRD4.

In the amino acid sequence of the first chimeric cysteine-rich region, in the amino acid sequence of the cysteine-rich region of wild-type DcR3, in addition to at least a portion of the CRD of wild-type DcR3, a portion other than the wild-type CRD is substituted with another peptide or protein. Sequences are also included. The portion other than CRD to be substituted with another peptide or protein may be selected from a region present between CRD1 and CRD2, a region present between CRD2 and CRD3, and a region present between CRD3 and CRD4. The moieties other than CRD substituted with other peptides or proteins may be one moiety or two or more moieties. For example, the amino acid sequence in which the amino acid sequence of the region between CRD1 and CRD2, the region between CRD2 and CRD3, or the region between CRD3 and CRD4 is substituted with another peptide or protein is also a first chimera It is included in the amino acid sequence of the cysteine-rich region.

In the cysteine-rich region of wild-type DcR3, one portion or two or more portions may be substituted with other peptides or proteins. When at least a portion of a CRD in the cysteine-rich region of wild-type DcR3 is substituted, the substituted portion may be all or part of one CRD or all or part of a plurality of CRDs, but may be affected by binding to LIGHT, TL1A and FasL It is more preferable that all or part of CRD2 and/or all or part of CRD3 region of the wild-type DcR3 involved are maintained, and all or part of other CRDs are substituted with other peptides or proteins.

As another peptide or protein to be substituted, any of natural or artificial peptides or proteins may be used. For example, at least a part of CRD derived from a protein other than DcR3 is exemplified. In a preferred embodiment, the other peptide or protein to be substituted is at least a part of the CRD of a TNF receptor superfamily (TNFRSF) molecule other than DcR3. That is, in a preferred embodiment, in the amino acid sequence of the cysteine-rich region of wild-type DcR3, in the first chimeric cysteine-rich region, at least a part of the cysteine-rich domain of wild-type DcR3 is the cysteine-rich domain of a TNFRSF molecule other than DcR3. and an amino acid sequence that is at least partially substituted. At least a portion of the CRD of the TNFRSF molecule may be selected from all or part of CRD1, all or part of CRD2, all or part of CRD3, and all or part of CRD4. Accordingly, the first chimeric cysteine-rich region is a substitution of a portion of CRD1 of wild-type DcR3, a portion of CRD1 of a TNFRSF molecule corresponding to a portion of CRD1 of wild-type DcR3, all of CRD1 of wild-type DcR3, CRD1 of a TNFRSF molecule Substitution in all of, a part of CRD2 of wild-type DcR3, substitution in a part corresponding to a part of CRD2 of wild-type DcR3 in CRD2 of a TNFRSF molecule, substitution of all of CRD2 of wild-type DcR3, substitution of all of CRD2 of TNFRSF molecule , a portion of CRD3 of wild-type DcR3, a substitution in a portion corresponding to a portion of CRD3 of wild-type DcR3 in CRD3 of a TNFRSF molecule, a substitution of all of CRD3 of wild-type DcR3, a substitution of all of CRD3 of a TNFRSF molecule, CRD4 of wild-type DcR3 may have one or more substitutions selected from a portion of CRD4 of a TNFRSF molecule in a portion corresponding to a portion of CRD4 of wild-type DcR3, and a substitution of all of CRD4 of wild-type DcR3 in all of CRD4 of a TNFRSF molecule. there is.

Examples of the first chimeric cysteine-rich region include, for example, a chimeric cysteine-rich region having CRD1 of wild-type DcR3 replaced with CRD1 of a TNFRSF molecule (in the chimeric cysteine-rich region, preferably other CRDs of wild-type DcR3) is maintained), a chimeric cysteine-rich region having a substitution of CRD4 of wild-type DcR3 with CRD4 of a TNFRSF molecule (in the chimeric cysteine-rich region, preferably other CRDs of wild-type DcR3 are maintained); or a chimeric cysteine-rich region (in the chimeric-type cysteine-rich region, preferably wild-type DcR3) having a wild-type DcR3 substitution of CRD1, a TNFRSF molecule with CRD1, and a wild-type DcR3 CRD4 of a TNFRSF molecule substitution. Other CRDs are maintained) and the like. Further, in addition to one or more of these substitutions, a substitution of a portion of CRD2 of wild-type DcR3 with a portion of CRD2 of a TNFRSF molecule corresponding to that portion, and/or of a portion of CRD3 of wild-type DcR3 corresponding to that portion Also included in the first chimeric cysteine-rich region is a chimeric cysteine-rich region further having a substitution with a portion of CRD3 of a TNFRSF molecule.

Human TNFRSF belongs to 29 receptors, any receptor has a CRD in the N-terminal extracellular domain, and it is typical that 6 Cys residues form 3 disulfide bonds with respect to 1 CRD, each receptor It has 1 to 4 CRDs according to [Trends Biochem Sci, 2002.27: p-19-26.].

Human TNFRSF, for example, DcR1 (TNFRSF10C, TRAIL-R3, LIT, TRID, CD263), DcR2 (TNFRSF10D, TRAIL-R4, TRUNDD, CD264), TNFR type I (TNFRSF1A, TNF-R, CD120a, TNFAR, TNF-R55, TNFR60), TNFR type II (TNFRSF1B, TNFBR, CD120b, TNFR80, p75, TNF-R75), LTBR (Lymphotoxin beta receptor), TNFRSF3, TNFR III, TNFCR, TNFR-RP, TNFR2 -RP), OX-40 (TNFRSF4, ACT35, TXGP1L, CD134), CD40 (TNFRSF5, Bp50, p50), Fas (Fas cell surface death receptor, TNFRSF6, CD95, APO-1, APT1) , FAS1), CD27 (TNFRSF7, S152, Tp55), CD30 (TNFRSF8, Ki-1), 4-1BB (TNFRSF9, CD137, ILA), DR4 (TNFRSF10A, Apo2, TRAILR-1, CD261), DR5 (TNFRSF10B, TRAIL-R2, KILLER, TRICK2A, TRICKB, CD262), RANK (TNFRSF11A, CD265, FEO), FN14 (TNFRSF12A, TweakR, CD266), TACI (TNFRSF13B, CD267, IGAD2), BAFFR (TNFRSF13C, CD268), HV14 , ATAR, TR2, LIGHTR, HVEA, CD270), NGFR (nerve growth factor receptor, TNFRSF16, p75NTR, CD271), BCMA (TNFRSF17, BCM, CD269, TNFRSF13A), GITR (TNFRSF18, AITR, CD357 ), TROY (TNFRSF19, TAJ-alpha, TAJ, TRADE), RELT (TNFRSF19L), DR6 (Death Receptor 6, TNFRSF21, CD358), DR3 (Death Receptor 3, TNFRSF25, TRAMP, WSL-1, LARD, WSL-LR, DDR3, TR3, APO-3), EDAR ( ectodysplasin A receptor, ED3, DL, ED5, EDA3, Edar, ED1R, EDA1R), EDAR2R (ectodysplasin A2 receptor, XEDAR, EDAA2R, EDA-A2R, TNFRSF27), or osteoprotegerin (OPG, TNFRSF11B, TR1, OCIF).

In the first chimeric-type cysteine-rich region, other peptides or proteins substituting at least a part of wild-type DcR3 are not limited, but OPG is particularly preferable among TNFRSF.

Although OPG is not limited to the origin, OPG derived from various eukaryotes is mentioned. For example, OPG derived from amphibians such as frogs, birds or mammals such as chickens, for example, primates including humans, ungulates such as pigs and cattle, and rodents including mice.

The cDNA sequence of human-derived OPG is shown in SEQ ID NO: 13, and the corresponding mRNA sequence is registered with GenBank (NCBI, USA) as accession number: NM_002546.3. The amino acid sequence of human-derived OPG is shown in SEQ ID NO: 14 and is registered with GenBank (NCBI, USA) as accession number: NP_002537.3.

The immature human OPG (SEQ ID NO: 14) has a signal peptide at the N-terminus. The mature human OPG is one in which the signal peptide is cleaved among the immature forms, and the amino acid sequence of the mature human OPG is, for example, SEQ ID NO: 16, and a DNA encoding the amino acid sequence of the mature human OPG. The nucleotide sequence of is shown, for example, in SEQ ID NO: 15. In the present invention, in the amino acid sequence of immature human OPG (SEQ ID NO: 14), the region 22 to 62 from the N-terminus is CRD1 (SEQ ID NO: 18), the region 65 to 105 is CRD2 (SEQ ID NO: 20), The 107th to 142th region is defined as CRD3 (SEQ ID NO: 22), and the 145th to 185th region is defined as CRD4 (SEQ ID NO: 24). The nucleotide sequences of DNA encoding the amino acid sequences of CRD1, CRD2, CRD3 and CRD4 of human OPG are shown, for example, in SEQ ID NOs: 17, 19, 21 and 23, respectively. In addition, the amino acid sequence of CRD has a plurality of definitions other than those described above, but any definition of the CRD amino acid sequence can be used for the DcR3 variant of the present invention using known information [UniProt O00300, GenBank NP_002537.3] .

In addition, polymorphisms and isoforms of genes are often found in genes encoding eukaryotic proteins. In the gene used in the present invention, a gene in which a mutation is generated in a nucleotide sequence or an amino acid sequence due to such a polymorphism is also included in the gene encoding the OPG of the present invention.

OPG inhibits bone destruction by osteoclasts by binding to RANKL and neutralizing its activity [J. Immunol., 2012, 189: p.245-252]. In addition, OPG also binds to TRAIL and inhibits apoptosis through TRAIL by neutralizing its activity [Am. J. Cancer. Res., 2012, 2: p.45-64]. Since there is a possibility that undesired activity or the like may be caused by neutralization of any ligand, it is preferable that the DcR3 variant of the present invention does not have neutralizing activity for either RANKL or TRAIL.

In one preferred embodiment, the first chimeric type cysteine-rich region contains the following amino acid sequences (a), (b), (c) or (d), or contains the amino acid sequence.

(a) In the amino acid sequence of the cysteine-rich region of wild-type DcR3, an amino acid sequence in which CRD1 of wild-type DcR3 is substituted with CRD1 of OPG (in the amino acid sequence, preferably other CRDs of wild-type DcR3 are maintained)

(b) an amino acid sequence in which CRD4 of wild-type DcR3 is substituted with CRD4 of OPG in the amino acid sequence of the cysteine-rich region of wild-type DcR3 (in the amino acid sequence, preferably other CRDs of wild-type DcR3 are maintained)

(c) in the amino acid sequence of the cysteine-rich region of wild-type DcR3, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG, and CRD4 of wild-type DcR3 is substituted with CRD4 of OPG (in the amino acid sequence, Preferably the other CRDs of wild-type DcR3 are maintained)

(d) in the amino acid sequence of (a), (b) or (c), a portion of CRD2 of wild-type DcR3 is substituted with a corresponding portion of CRD2 of OPG, and/or a portion of CRD3 of wild-type DcR3 is Amino acid sequence substituted with the corresponding portion of CRD3 of OPG (in the corresponding amino acid sequence, preferably other CRDs of wild-type DcR3 are maintained)

As the amino acid sequence of (d), for example, in the amino acid sequence of (a), (b) or (c), the 103rd to 123rd positions from the N-terminus correspond to the corresponding amino acid sequence of the OPG CRD. and partially substituted amino acid sequences. In this amino acid sequence, the amino acid sequence comprising the 18th to 36th portion of CRD3 of wild-type DcR3 and 2 amino acid residues following the C-terminal side of the CRD3 amino acid sequence is substituted with a portion of the OPG corresponding to the amino acid sequence there is.

Specific examples of the amino acid sequence of (a) include an amino acid sequence containing the amino acid positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 26 or 50, and a specific example of the amino acid sequence of (b) above , an amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence shown in SEQ ID NO: 28 or 52, and specific examples of the amino acid sequence of (c) include the amino acid sequence shown in SEQ ID NO: 30 or 54 Among them, an amino acid sequence containing the amino acid at positions 1-164 from the N-terminus is mentioned, and as a specific example of the amino acid sequence of (d), the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 32 or 56. and amino acid sequences comprising

SEQ ID NO: 26 is the amino acid sequence of chimeric B-HBD (in wild-type DcR3 (SEQ ID NO: 4), a DcR3 variant in which CRD1 is substituted with CRD1 of OPG), SEQ ID NO: 28 is chimeric C-HBD (wild-type DcR3) In (SEQ ID NO: 4), the amino acid sequence of the DcR3 variant in which CRD4 was substituted with CRD4 of OPG), SEQ ID NO: 30, is the chimeric A-HBD (wild-type DcR3 (SEQ ID NO: 4), CDR1 and CDR4 are respectively OPG The amino acid sequence, SEQ ID NO: 32 of the DcR3 variant substituted with CDR1 and CDR4 of The amino acid sequence, SEQ ID NO: 50 of the DcR3 variant in which the amino acid sequence containing The amino acid sequence of the DcR3 variant with the binding region deleted), SEQ ID NO: 52, in chimeric C (wild type DcR3 (SEQ ID NO: 4), substituted CRD4 with CRD4 of OPG, and DcR3 with the heparan sulfate binding region deleted) The amino acid sequence of the variant), SEQ ID NO: 54, is chimera A (in wild-type DcR3 (SEQ ID NO: 4), CDR1 and CDR4 are substituted with OPG CDR1 and CDR4, respectively, and DcR3 with a heparan sulfate binding region deleted) The amino acid sequence of the variant), SEQ ID NO: 56, is 103-123OPG (in chimera A, DcR3 in which the amino acid sequence containing the 18th to 36th portions of CRD3 and 2 amino acids on the C-terminal side thereof is substituted with human OPG) variant) of the amino acid sequence.

In a preferred embodiment, as a DcR3 variant comprising a first chimeric cysteine-rich region, a DcR3 variant having binding activity to at least one or more of LIGHT, TL1A and FasL, LIGHT, binding to all of TL1A and FasL DcR3 variant having activity, without binding activity to FasL, and also having binding activity to any one of LIGHT and TL1A, DcR3 variant having no binding activity to FasL, and binding to LIGHT and TL1A DcR3 variants having activity may be mentioned.

In the present invention, "having binding activity to a ligand" means that the binding activity of the DcR3 variant comprising the first chimeric cysteine-rich region to the ligand is compared with that of wild-type DcR3 to the ligand. It is equivalent and is used in the meaning including the thing which is not significantly reduced, and the thing which is significantly enhanced compared with the binding activity of wild-type DcR3 with respect to the said ligand. For example, when measured by surface plasmon resonance (SPR), when the dissociation constant (K _D ) value of the DcR3 variant is less than three times that of wild-type DcR3, it can be judged that it has binding activity to the ligand. can

In the present invention, the expression "doesn't have binding activity to the ligand" of the variant DcR3 means that the binding activity of the variant DcR3 comprising the first chimeric cysteine-rich region to the ligand is not detected, and Compared with the binding activity of wild-type DcR3 to the ligand, it is used in a meaning including a significantly lowered one. For example, when measured by the SPR method, when the K _D value of the DcR3 variant is greater than 3 times compared to the wild-type DcR3, or the Rmax of the DcR3 variant is less than 5, the binding activity to the ligand is preferentially lowered It can be determined that the DcR3 variant does not have binding activity to the ligand.

In one particularly preferred embodiment, the DcR3 variant having the first chimeric type cysteine-rich region includes a DcR3 variant having reduced binding to FasL. "FasL binding lowering variant" is a DcR variant comprising a chimeric cysteine-rich region that does not have binding activity to FasL, and has binding activity to any one or more of LIGHT and TL1A, or binding to FasL It does not have activity, and also refers to a DcR3 variant comprising a chimeric cysteine-rich region having binding activity to LIGHT and TL1A.

In a preferred embodiment, the DcR3 variant of the present invention is a DcR3 variant having neutralizing activity with respect to at least one of LIGHT, TL1A and FasL, LIGHT, a DcR3 variant having neutralizing activity with respect to all of TL1A and FasL, It is a DcR3 variant that does not have neutralizing activity with respect to FasL and has neutralizing activity with respect to any one of LIGHT and TL1A, or a DcR3 variant that does not have neutralizing activity with respect to FasL and has neutralizing activity with respect to LIGHT and TL1A. .

In the present invention, the expression "neutralizing activity for a certain ligand" means that the ligand binds to a DcR3 variant, thereby inhibiting binding of the ligand to a receptor on the cell membrane surface, and a receptor on the cell membrane surface of the ligand Cell functions caused by binding of the ligand to a receptor on the cell membrane surface by inhibiting binding to the ligand, that is, the biological activity of the ligand (eg, biological activity such as cytokine production, proliferation enhancement, induction of apoptosis, etc.) includes impeding

In the present invention, the expression "having a neutralizing activity" of a DcR3 variant means that the neutralizing activity of the DcR3 variant for the ligand is compared with the neutralizing activity of wild-type DcR3 for the ligand, and there is no significant difference; and Compared with the neutralizing activity of wild-type DcR3 with respect to the ligand, it is used in a meaning including significantly enhanced.

In the present invention, the expression "does not have neutralizing activity" of the modified DcR3 variant means that the neutralizing activity of the DcR3 variant with respect to the ligand is significantly reduced compared to that of the wild-type DcR3 for the ligand. used in the sense of including

In one particularly preferred embodiment, the modified DcR3 in the present invention is a modified DcR3 having reduced binding to FasL. "FasL binding lowering variant" does not have neutralizing activity with respect to FasL, and DcR3 variant having neutralizing activity with respect to any one or more of LIGHT and TL1A, or FasL It does not have neutralizing activity, and LIGHT and It refers to a DcR3 variant having neutralizing activity with respect to TL1A.

2-1-2. second chimeric cysteine rich region

The second chimeric-type cysteine-rich region is one in which 1 to 30 amino acids are deleted, substituted, inserted or added in the amino acid sequence of the first chimeric-type cysteine-rich region.

As a method for obtaining a polypeptide having an amino acid sequence in which one or more amino acids are deleted, substituted, inserted or added in the amino acid sequence of the first chimeric cysteine-rich region, site-directed mutagenesis [Molecular Cloning, A Laboratory Manual, Second Edition] , Cold Spring Harbor Laboratory Press (1989), Current Protocols inmolecular Biology, John Wiley & Sons (1987-1997), Nucleic Acids Research, 10, 6487 (1982), Proc. Natl. Acad. Sci. USA., 79, 6409, (1982), Gene, 34, 315 (1985), Proc. Natl. Acad. Sci. USA., 82, 488 (1985)].

Mutations (modifications) applied to the first chimeric cysteine-rich region include natural mutations and artificial amino acid substitutions, deletions, insertions, or additions. As the amino acid sequence of the second chimeric cysteine-rich region, the first In the amino acid sequence of the chimeric cysteine-rich region, 1 or 2 or more, preferably 1 to 30, more preferably 1 to 10, still more preferably 1 to 5, still more preferably An amino acid sequence in which 1 to 3 amino acids are substituted, deleted, inserted or added, or an amino acid sequence of the first chimeric type cysteine-rich region is 80% or more, preferably 85% or more, more preferably 90% or more, e.g. For example, an amino acid sequence having 93% or more, 95% or more, 97% or more, 98% or more, or 99% or more identity. As a method of describing an amino acid substitution, for example, when the Asn at position 131 from the N-terminus of the amino acid sequence to be substituted is substituted with Ser, it can be expressed as N131S.

In one preferred embodiment, the second chimeric cysteine-rich region includes the following amino acid sequence (e) or includes the amino acid sequence.

(e) in the amino acid sequence of (a), (b), (c) or (d), 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence.

Examples of the mutation (modification) applied to the first chimeric cysteine-rich region include addition or deletion of a sugar chain addition site. By addition or deletion of a sugar chain addition site to the first chimeric type cysteine-rich region, biological activity or properties thereof, in vivo dynamics such as blood half-life, or physical or chemical properties such as protein stability of the DcR3 variant of the present invention can be controlled.

Sugar chain addition generally refers to N-glycosidic bonding of a sugar chain to an asparagine residue of a peptide or protein and/or O-glycosidic bonding of a sugar chain to a serine or threonine residue. Examples of the O-type sugar chain added to the DcR3 variant include core 1, core 2, and the like, and examples of the N-type sugar chain include hymannose, hybrid, or complex sugar chains, but are preferably complex sugar chains.

As a mutation (modification) applied to the first chimeric type cysteine-rich region, for example, at least one or more amino acids in the amino acid sequence of the first chimeric type cysteine-rich region are replaced by an N-glycosidic bond or an O-glycosidic bond. Examples include adding a sugar chain by substituting an amino acid capable of adding a sugar chain, and it is particularly preferable to add an N-glycoside-linked sugar chain.

Also, for example, in the amino acid sequence of the chimeric cysteine-rich region, at least one or more, preferably two or more, amino acids involved in N-glycosidic bonds are substituted with other amino acids and sugar chains are removed according to the present invention. included in In general, when a peptide or protein is expressed using yeast, insect cells or mammalian cells, the Asn-X-Thr/Ser (where X is any amino acid residue other than Pro) no sequence is recognized and sugar chain of N-glycosidic bonds. For example, by substituting another amino acid for Asn, Ser, or Thr in the Asn-X-Thr/Ser sequence present in the DcR3 variant, the N-glycoside-linked sugar chain can be removed.

As the second chimeric-type cysteine-rich region, in order to reduce the aggregate of the DcR3 variant of the present invention, in particular, the amino acid sequence of the first chimeric-type cysteine-rich region (for example, in SEQ ID NO: 30, 1 to 164th from the N-terminus) A chimeric cysteine-rich region having a sugar chain N-glycosidically bound to Asn at position 157 from the N-terminus of the amino acid sequence (amino acid sequence containing amino acids) is preferred.

In one preferred embodiment, the amino acid sequence of the second chimeric cysteine-rich region from which the sugar chain addition site has been removed (one form of the amino acid sequence of (e) above) is:

(f) the amino acid sequence of (b), (c) or (d) (for example, the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56 Substitution of Asn at positions 131 and 144 from the N-terminus of the amino acid sequence comprising a

(g) the amino acid sequence of (b), (c) or (d) (eg, amino acids 1 to 164 from the N-terminus in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56 Substitution of Asn at positions 131, 144 and 157 from the N-terminus of the amino acid sequence comprising a

(h) the amino acid sequence of (b), (c) or (d) (for example, the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56 Substitution with other amino acids of Thr at position 133 and Ser at position 146 from the N-terminus of the amino acid sequence comprising

(i) the amino acid sequence of (b), (c) or (d) (for example, amino acids 1 to 164 from the N-terminus in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56 from the N-terminus of the amino acid sequence comprising

has a substitution selected from

In a more preferred embodiment, the amino acid sequence from which the sugar chain addition site has been removed (one form of the amino acid sequence of (e) above) is:

(f ') the amino acid sequence of (b), (c) or (d) (for example, in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56, 1 to 164th from the N-terminus Substitution of Asn at positions 131 and 144 from the N-terminus of an amino acid sequence comprising an amino acid) with Ser;

(g ') the amino acid sequence of (b), (c) or (d) (for example, in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56, 1 to 164th from the N-terminus Substitution of Asn at positions 131, 144 and 157 from the N-terminus of the amino acid sequence comprising an amino acid) to Ser;

(h ') the amino acid sequence of (b), (c) or (d) (for example, in the amino acid sequence shown in SEQ ID NOs: 28, 30, 32, 52, 54 or 56, 1 to 164th from the N-terminus Thr at position 133 from the N-terminus of the amino acid sequence comprising an amino acid) and Ser at position 146 with Ala, and

(i') the amino acid sequence of (b), (c) or (d) (for example, in the amino acid sequence shown in SEQ ID NO: 28, 30, 32, 52, 54 or 56, 1 to 164th from the N-terminus amino acid sequence comprising an amino acid) from the N-terminus of Thr at position 133, Ser at position 146, and Thr at position 159 with Ala

has a substitution selected from

As the amino acid sequence having the substitution of (f') above, for example, in the amino acid sequence shown in SEQ ID NO: 30 or SEQ ID NO: 54, in the amino acid sequence containing the amino acids 1 to 164 from the N terminus, 131 from the N terminus and an amino acid sequence in which the Asn at positions 144 and Ser is substituted (N131S/N144S) (an amino acid sequence containing amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 34 or SEQ ID NO: 58) can

In addition, SEQ ID NO: 34 is the amino acid sequence of N131S/N144S-HBD (in chimeric A-HBD, the DcR3 variant in which Asn at positions 131 and 144 from the N-terminus is substituted with Ser), SEQ ID NO: 58 is N131S/ The amino acid sequence of N144S (in chimera A, a DcR3 variant in which Asn at positions 131 and 144 from the N-terminus is substituted with Ser) is shown.

As the amino acid sequence having the substitution of (h') above, for example, in the amino acid sequence shown in SEQ ID NO: 30 or SEQ ID NO: 54, in the amino acid sequence containing the amino acids 1 to 164 from the N terminus, 133 from the N terminus Amino acid sequence in which Thr at position 146 and Ser at position 146 are substituted with Ala (T133A/S146A) (an amino acid sequence containing amino acids 1 to 164 from the N-terminus in the amino acid sequence shown in SEQ ID NO: 36 or SEQ ID NO: 60), etc. can be heard

In addition, SEQ ID NO: 36 is the amino acid sequence of T133A/S146A-HBD (in chimeric A-HBD, the DcR3 variant in which Thr at position 133 and Ser at position 146 from the N-terminus are substituted with Ala), SEQ ID NO: 60 is, The amino acid sequence of T133A/S146A (in chimera A, the DcR3 variant in which Thr at position 133 and Ser at position 146 from the N-terminus are substituted with Ala) is shown.

As the amino acid sequence having the substitution of (g') above, for example, in the amino acid sequence containing the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30 or SEQ ID NO: 54, the amino acid sequence at position 131 from the N-terminus , an amino acid sequence in which Asn at positions 144 and 157 is substituted with Ser (N131S/N144S/N157S) (in the amino acid sequence set forth in SEQ ID NO: 38 or SEQ ID NO: 62, an amino acid sequence containing amino acids 1 to 164 from the N-terminus ) and the like.

In addition, SEQ ID NO: 38 is the amino acid sequence of N131S/N144S/N157S-HBD (in chimeric A-HBD, the DcR3 variant in which Asn at positions 131, 144, and 157 from the N-terminus is substituted with Ser), SEQ ID NO: 62 shows the amino acid sequence of N131S/N144S/N157S (in chimera A, a DcR3 variant in which Asn at positions 131, 144, and 157 from the N-terminus is substituted with Ser).

As the amino acid sequence having the substitution of (i'), for example, in the amino acid sequence shown in SEQ ID NO: 30 or SEQ ID NO: 54, in the amino acid sequence containing the amino acids 1 to 164 from the N-terminus, 133 from the N-terminus An amino acid sequence in which Thr at position 146, Ser at position 146 and Thr at position 159 are substituted with Ala (T133A/S146A/T159A) (in the amino acid sequence shown in SEQ ID NO: 40 or SEQ ID NO: 64, amino acids 1 to 164 from the N-terminus) amino acid sequence comprising a) and the like.

SEQ ID NO: 40 is the amino acid of T133A/S146A/T159A-HBD (in chimeric A-HBD, DcR3 variant in which Thr at position 133, Ser at position 146, and Thr at position 159 from the N-terminus are substituted with Ala) The sequence, SEQ ID NO: 64, shows the amino acid sequence of T133A/S146A/T159A (in chimera A, a DcR3 variant in which Thr at 133, Ser at position 146, and Thr at position 159 from the N-terminus are substituted with Ala).

In the mutation (formula) applied to the first chimeric cysteine-rich region, for example, no mutation is added to CRD2 and CRD3 of the first chimeric cysteine-rich region involved in binding to LIGHT, TL1A and FasL, 1 By introducing a mutation into CRD1 and/or CRD4 of the chimeric cysteine-rich region, a DcR3 variant containing a chimeric cysteine-rich region in which the binding activity between LIGHT, TL1A and FasL and the chimeric cysteine-rich region is not reduced can be obtained. can On the other hand, by introducing a mutation in CRD2 or/and CRD3 of the first chimeric cysteine-rich region involved in binding to LIGHT, TL1A or FasL, the binding activity between LIGHT, TL1A or FasL and the chimeric cysteine-rich region is changed. A DcR3 variant containing a cysteine-rich region can be obtained. That is, by introducing the above-described mutation, it is possible to obtain a DcR3 variant comprising a chimeric cysteine-rich region having desirable LIGHT, TL1A or FasL binding properties.

In a preferred embodiment, as the DcR3 variant comprising a second chimeric cysteine-rich region, a DcR3 variant comprising a chimeric cysteine-rich region having binding activity to at least one or more of LIGHT, TL1A and FasL, LIGHT, A DcR3 variant comprising a chimeric cysteine-rich region having binding activity to both TL1A and FasL, a chimeric cysteine-rich region having no binding activity to FasL and having binding activity to any one of LIGHT and TL1A and a DcR3 variant comprising a, or a DcR3 variant comprising a chimeric cysteine-rich region that does not have binding activity to FasL, and has binding activity to LIGHT and TL1A. The meanings of the expressions "have binding activity to a ligand" and "does not have binding activity to a ligand" of the modified DcR3 of the present invention are as described above.

In a particularly preferred embodiment, examples of the DcR3 variant containing the second chimeric type cysteine-rich region include a DcR3 variant with reduced binding to FasL. "FasL binding lowering variant" is a DcR3 variant comprising a chimeric cysteine-rich region that has no binding activity to FasL, and has binding activity to any one or more of LIGHT and TL1A, or FasL binding It does not have activity, and also refers to a DcR3 variant comprising a chimeric cysteine-rich region having binding activity to LIGHT and TL1A.

In a preferred embodiment, as the DcR3 variant comprising a second chimeric cysteine-rich region, a DcR3 variant comprising a chimeric cysteine-rich region having neutralizing activity against at least one or more of LIGHT, TL1A and FasL, LIGHT, DcR3 variant comprising a chimeric cysteine-rich region having neutralizing activity against all of TL1A and FasL, a chimeric cysteine-rich region that does not have neutralizing activity against FasL and has neutralizing activity against either LIGHT or TL1A and a DcR3 variant comprising a DcR3 variant comprising a chimeric cysteine-rich region that does not have neutralizing activity against FasL, and has neutralizing activity against LIGHT and TL1A, and the like. The meanings of the expressions "having neutralizing activity with respect to a ligand" and "having no neutralizing activity with respect to a ligand" of the modified DcR3 of the present invention are as described above.

In one particularly preferred embodiment, as the DcR3 variant containing the second chimeric type cysteine-rich region, a DcR3 variant having reduced neutralizing activity for FasL is exemplified. "FasL binding lowering variant" does not have neutralizing activity with respect to FasL, and also has neutralizing activity with respect to any one or more of LIGHT and TL1A DcR3 variant comprising a chimeric cysteine-rich region, or neutralizing with respect to FasL It has no activity, and also refers to a DcR3 variant comprising a chimeric cysteine-rich region having neutralizing activity with respect to LIGHT and TL1A.

For example, in a DcR3 variant including a chimeric cysteine-rich region, which is a variant that decreases FasL binding, the binding site of DcR3 or DcR3 variant, which is estimated from crystal structure analysis, etc., to each ligand is substituted with Ala or another amino acid. It can be obtained by constructing a modified form and measuring the binding activity and neutralizing activity to LIGHT, TL1A or FasL ligand. In addition, a gene library in which the periphery of the ligand binding site of DcR3 or DcR3 variant is randomly converted into other amino acids is prepared and displayed on phage, yeast, mammalian cells, etc., binding activity and neutralizing activity to LIGHT, TL1A or FasL ligand It can also be obtained by screening as an index.

As the amino acid sequence of the chimeric cysteine-rich region (one form of the amino acid sequence of (e) above) contained in the FasL binding lowering variant, for example, (a), (b), (c) or (d) and amino acid sequences having substitution of one or two or more amino acids selected from the group consisting of Glu at position 57, Arg at position 58, and Arg at position 60 from the N-terminus of the amino acid sequence of

The other amino acids substituted for Glu at position 57 are not particularly limited, and 20 kinds of amino acids (Glu, Ala, Asp, Lys, Leu, Cys, Phe, Gly, His, Ile, Met, Asn, Pro, Gln, Arg) , Ser, Thr, Val, Trp, Tyr) can be appropriately selected from 19 types of amino acids except for Glu, but is preferably selected from Lys, Leu, Arg, Val, Ala, Phe, His, He and Met, More preferably, it is selected from Lys, Leu, Arg and Val, even more preferably selected from Lys, Arg and Val, and still more preferably selected from Lys and Arg.

The other amino acid substituted for Arg at position 58 is not particularly limited, and can be appropriately selected from 19 types of amino acids excluding Arg among 20 types of amino acids. It is more preferable to select from

The other amino acid substituted for Arg at position 60 is not particularly limited and can be appropriately selected from 19 types of amino acids excluding Arg among 20 types of amino acids, but is preferably Lys.

In a preferred embodiment, one or two or more amino acids selected from the group consisting of Glu at position 57, Arg at position 58, and Arg at position 60 is one amino acid including Glu at position 57.

In another preferred embodiment, one or two or more amino acids selected from the group consisting of Glu at position 57, Arg at position 58, and Arg at position 60 are two amino acids including Glu at position 57 and Arg at position 58. is an amino acid. In this aspect, it is preferable to combine the substitution of Lys, Leu, Arg, Val, Ala, Phe, His, Ile or Met of Glu at position 57 with the substitution of Asp, Glu or Thr of Arg at position 58. and a substitution in Lys, Leu, Arg or Val of the 57th Glu and a substitution in Asp or Glu of the 58th Arg are more preferably combined, and the substitution in Lys or Arg of the 57th Glu and , it is still more preferable to combine the substitution in Asp or Glu of Arg at the 58th position.

The second chimeric type cysteine-rich region may have substitution of one or two or more amino acids other than Glu at position 57, Arg at position 58, and Arg at position 60 with other amino acids. As amino acids other than Glu at position 57, Arg at position 58, and Arg at position 60, for example, Trp at position 53 from the N-terminus of the amino acid sequence of (a), (b), (c) or (d) above; Asn at the 54th position, Tyr at the 55th position, Leu at the 56th position, and the like are mentioned.

The other amino acid substituted for Trp at position 53 is not particularly limited and can be appropriately selected from 19 types of amino acids excluding Trp from among 20 types of amino acids, but is preferably selected from Asp and Asn. Substitution of Trp at position 53 with another amino acid can be combined with substitution of Glu at position 57 with another amino acid, for example.

The other amino acid substituted with Asn at position 54 is not particularly limited and can be appropriately selected from 19 types of amino acids excluding Asn from among 20 types of amino acids, but is preferably Asp. Substitution of Asn at position 54 in another amino acid can be combined with substitution of Glu at position 57 in another amino acid, for example.

The other amino acid substituted with Tyr at position 55 is not particularly limited, and can be appropriately selected from 19 types of amino acids excluding Tyr among 20 types of amino acids, but is preferably selected from Thr, Asp, Gin and Glu. Substitution of Tyr at position 55 with another amino acid can be combined with substitution of Glu at position 57 with another amino acid, for example.

The other amino acid substituted for Leu at position 56 is not particularly limited, and can be appropriately selected from 19 amino acids excluding Leu among 20 amino acids, and selected from Asp, Gin, Thr, Glu, Gly, Asn and Pro. It is preferable to do Substitution of Leu at position 56 with another amino acid can be combined with substitution of Glu at position 57 with another amino acid, for example.

1 or 2 or more amino acids selected from the group consisting of Glu at position 57, Arg at position 58, and Arg at position 60 from the N-terminus of the amino acid sequence of (a), (b), (c) or (d) Of, as an amino acid sequence having a substitution with another amino acid, specifically,

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Lys (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 42) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Lys (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 66) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Leu (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 44) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Leu (N-terminal in the amino acid sequence set forth in SEQ ID NO: 68) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence containing the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Arg at position 60 from the N-terminus is substituted with Lys (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 46) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Arg at position 60 from the N-terminus is substituted with Lys (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 70) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, an amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Arg;

In the amino acid sequence containing the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Arg (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 180) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, an amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Val;

In the amino acid sequence containing the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Val (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 182) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Ala;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Ala (N-terminal in the amino acid sequence set forth in SEQ ID NO: 270) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Phe;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Phe (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 272) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, an amino acid sequence in which Glu at position 57 from the N-terminus is substituted with His;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, an amino acid sequence in which Glu at position 57 from the N-terminus is substituted with His (N-terminal in the amino acid sequence set forth in SEQ ID NO: 274) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Ile;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Ile (N-terminal in the amino acid sequence set forth in SEQ ID NO: 276) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Met;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Met (the N-terminus in the amino acid sequence set forth in SEQ ID NO: 278) an amino acid sequence comprising the amino acid at positions 1 to 164 from

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30, Glu at position 57 from the N-terminus is substituted with Lys, and Arg at position 58 is substituted with Asp;

In the amino acid sequence comprising the amino acid positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from N-terminus is substituted with Lys and Arg at position 58 is substituted with Asp (SEQ ID NO: An amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus among the amino acid sequences described in 184);

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Lys and Arg at position 58 is substituted with Glu;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from N-terminus is substituted with Lys and Arg at position 58 with Glu (SEQ ID NO: an amino acid sequence comprising the amino acids 1 to 164 from the N-terminus among the amino acid sequences described in 186);

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, Glu at position 57 from the N-terminus is substituted with Arg, and Arg at position 58 is substituted with Asp;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 54, Glu at position 57 from the N-terminus is substituted with Arg, and Arg at position 58 is substituted with Asp (SEQ ID NO: of the amino acid sequence described in 188, an amino acid sequence comprising the amino acids 1 to 164 from the N-terminus);

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, the amino acid sequence in which Glu at position 57 from the N-terminus is substituted with Lys and Arg at position 58 is substituted with Thr;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, Glu at position 57 from N-terminus is substituted with Lys, and Arg at position 58 is substituted with Thr (SEQ ID NO: of the amino acid sequence described in 280, the amino acid sequence comprising the amino acids 1 to 164 from the N-terminus);

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, Glu at position 57 from the N-terminus is substituted with Leu, and Arg at position 58 is substituted with Glu;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 54, Glu at position 57 from N-terminus is substituted with Leu, and Arg at position 58 is substituted with Glu (SEQ ID NO: an amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus among the amino acid sequences described in 282);

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, Glu at position 57 from the N-terminus is substituted with Val, and Arg at position 58 is substituted with Thr;

In the amino acid sequence comprising the amino acid positions 1 to 164 from the N-terminus among the amino acid sequence set forth in SEQ ID NO: 54, the amino acid sequence in which Glu at position 57 from N-terminus is substituted with Val and Arg at position 58 is substituted with Thr (SEQ ID NO: An amino acid sequence comprising the amino acids 1 to 164 from the N-terminus among the amino acid sequences described in 284);

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus of the amino acid sequence set forth in SEQ ID NO: 30, Glu at position 57 from the N-terminus is substituted with Val, and Arg at position 58 is substituted with TGlu;

In the amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus among the amino acid sequence set forth in SEQ ID NO: 54, Glu at position 57 from N-terminus is substituted with Val and Arg at position 58 is substituted with Glu (SEQ ID NO: 286, the amino acid sequence comprising the amino acids 1 to 164 from the N-terminus)

and the like.

SEQ ID NO: 42 is the amino acid sequence of chimeric A-E57K-HBD (in chimeric A-HBD, the DcR3 variant in which Glu at the 57th position from the N-terminus is substituted with Lys), SEQ ID NO: 44 is chimeric A-E57L The amino acid sequence of -HBD (DcR3 variant in which Glu at position 57 from the N-terminus was substituted with Leu in chimeric A-HBD), SEQ ID NO: 46 is chimeric A-R60K-HBD (in chimeric A-HBD, N The amino acid sequence of the DcR3 variant in which Arg at position 60 from the terminal was substituted with Lys), SEQ ID NO: 66, is chimera A-E57K (DcR3 variant in which Glu at position 57 from the N-terminal is substituted with Lys in chimera A) The amino acid sequence of SEQ ID NO: 68 is the amino acid sequence of chimera A-E57L (in chimera A, the DcR3 variant in which Glu at position 57 from the N-terminus is substituted with Leu), SEQ ID NO: 70 is chimera A-R60K (chimera) In A, the amino acid sequence of the DcR3 variant in which Arg at position 60 from the N-terminus was substituted with Lys), SEQ ID NO: 180, is chimera A-E57R (In chimera A, Glu at position 57 from N-terminus is substituted with Arg) One DcR3 variant) amino acid sequence, SEQ ID NO: 182 is the amino acid sequence of chimera A-E57V (in chimera A, a DcR3 variant in which Glu at the 57th position from the N-terminus is substituted with Val), SEQ ID NO: 184 is chimera A The amino acid sequence of -E57K_R58D (in chimera A, the DcR3 variant in which Glu at position 57 from the N-terminus was substituted with Lys and Arg at position 58 with Asp), SEQ ID NO: 186, is chimera A-E57K_R58E (in chimera A) , the amino acid sequence, SEQ ID NO: 188 of the DcR3 variant in which Glu at position 57 from N-terminus was substituted with Lys and Arg at position 58 with Glu) is chimera A-E57R_R58D (in chimera A, at position 57 from N-terminus) Glu to Arg, DcR3 variant in which Arg at position 58 is substituted with Asp) The amino acid sequence, SEQ ID NO: 270 is the amino acid sequence of chimera A-E57A (in chimera A, the DcR3 variant in which Glu at the 57th position from the N-terminus is substituted with Ala), SEQ ID NO: 272 is chimera A-E57F (chimera A In the amino acid sequence, SEQ ID NO: 274 of the DcR3 variant in which the Glu at the 57th position from the N-terminus was substituted with Phe), the chimera A-E57H (in the chimera A, the Glu at the 57th position from the N-terminus was substituted with His The amino acid sequence of the DcR3 variant), SEQ ID NO: 276, is the amino acid sequence of chimera A-E57I (in chimera A, the DcR3 variant in which the 57th Glu from the N-terminus is substituted with Ile), SEQ ID NO: 278 is the chimera A The amino acid sequence, SEQ ID NO: 280 of the amino acid sequence of -E57M (DcR3 variant in which Glu at position 57 from the N-terminus was substituted with Met in chimera A) is chimera A-E57K_R58T (in chimera A, Glu at position 57 from N-terminus in A) The amino acid sequence, SEQ ID NO: 282 of the DcR3 variant substituted with Lys and Arg at position 58 with Thr) is chimera A-E57K_R58T (in chimera A, Glu at position 57 from the N-terminus is replaced with Lys, The amino acid sequence of the DcR3 variant in which Arg was substituted with Thr), SEQ ID NO: 284, is chimera A-E57V_R58T (in chimera A, DcR3 in which Glu at position 57 from the N-terminus is substituted with Val, and Arg at position 58 is substituted with Thr The amino acid sequence of the variant), SEQ ID NO: 286, is the amino acid sequence of chimera A-E57V_R58E (in chimera A, the DcR3 variant in which Glu at position 57 from the N-terminus was substituted with Val, and Arg at position 58 was substituted with Glu) indicates.

2-2. other areas

The DcR3 variant of the present invention may or may not include one or two or more other regions bound to the C-terminal side of the first or second chimeric cysteine-rich region. Other regions include, for example, part or all of the region between CRD4 and HBD in wild-type DcR3, and a part of the region following the C-terminal end of the CRD4 amino acid sequence in TNF receptor superfamily (TNFRSF) molecules other than DcR3. or all, part or all of HBD of wild-type DcR3, and the like. The expression "other regions bound to the C-terminal side of the first or second chimeric-type cysteine-rich region" means that when the other region is directly bound to the C-terminus of the first or second chimeric-type cysteine-rich region, and other regions are used in the meaning including the case where the C terminus of the first or second chimeric type cysteine-rich region is bound through another region.

In one embodiment, the DcR3 variant of the present invention is another region bound to the C-terminal side of the first or second chimeric-type cysteine-rich region, and part or all of a region present between CRD4 and HBD in wild-type DcR3. includes In this aspect, it is preferable that the other regions are directly bonded to the C terminus of the first or second chimeric cysteine-rich region.

In another embodiment, the DcR3 variant of the present invention is another region bound to the C-terminal side of the first or second chimeric type cysteine-rich region, and is an amino acid of CRD4 in a TNF receptor superfamily (TNFRSF) molecule other than DcR3. contains some or all of the region continuing at the C-terminus of the sequence. In this aspect, it is preferable that the other regions are directly bonded to the C terminus of the first or second chimeric cysteine-rich region. The description regarding TNFRSF is the same as the above. TNFRSF is preferably OPG.

In the first or second chimeric cysteine-rich region (eg, the first chimeric cysteine-rich region comprising the amino acid sequences of (b) to (d) above), CRD4 of wild-type DcR3 is substituted with CRD4 of OPG In this case, the DcR3 variant of the present invention preferably contains a plurality of amino acid residues following the C terminus of the CRD4 amino acid sequence of OPG as another region bound to the C-terminus of the chimeric cysteine-rich region. That is, when CR4 of wild-type DcR3 is substituted with CRD4 of OPG, a plurality of amino acid residues following the C terminus of the amino acid sequence of CRD4 of wild type DcR3 are also substituted with a plurality of amino acid residues following the C terminus of CRD4 amino acid sequence of OPG It is preferable to be As the plurality of amino acid residues following the C-terminus of the CRD4 amino acid sequence of OPG, for example, the 186th to 194th amino acid residues in the OPG amino acid sequence (SEQ ID NO: 14) are preferred, but at the C-terminus of CRD4 of wild-type DcR3 The number of consecutive amino acid residues and the number of amino acid residues to be substituted can be appropriately adjusted. The number of the plurality of amino acid residues following the C terminus of the CRD4 amino acid sequence of OPG is usually 1 to 12, preferably 1 to 10, more preferably 1 to 9, 1 to 6, or 1 to it is three In addition, in the first or second chimeric cysteine-rich region (eg, the first chimeric cysteine-rich region comprising the amino acid sequences of (a) and (d) above), when CRD4 is derived from wild-type DcR3, It is preferable that the amino acid sequence following the C terminus of the CRD4 amino acid sequence of wild-type DcR3 binds to the C-terminus of the amino acid sequence of CRD4 in the first chimeric cysteine-rich region. Examples of such an amino acid sequence include amino acids 194 to 195 of SEQ ID NO: 2, but the number of amino acid residues constituting the amino acid sequence continuing to the C terminus of the amino acid sequence of CRD4 of wild-type DcR3 can be appropriately adjusted. there is. The number of amino acid residues constituting the amino acid sequence continuing to the C-terminus of CRD4 of wild-type DcR3 is usually 1 to 10, preferably 1 to 5, more preferably 1 to 3, still more preferably 1 to It is two.

In another embodiment, the DcR3 variant of the present invention is another region bound to the C-terminal side of the first or second chimeric-type cysteine-rich region, and contains a part or all of HBD of wild-type DcR3. In this embodiment, the other regions may be directly bound to the C-terminus of the first or second chimeric-type cysteine-rich region, or CRD4 in wild-type DcR3 to the C-terminus of the first or second chimeric-type cysteine-rich region. and HBD, or through part or all of the region following the C terminus of the CRD4 amino acid sequence in a TNFRSF molecule other than DcR3.

The DcR3 variant of the present invention is another region bound to the C-terminal side of the first or second chimeric type cysteine-rich region, does not include HBD, and is at positions 186 to 194 in the amino acid sequence of OPG (SEQ ID NO: 14) It is preferred to include an amino acid residue of

3. DcR3 variants containing sugar chains

The DcR3 variant of the present invention includes a DcR3 variant including at least one sugar chain. As long as at least one sugar chain is bound to the cysteine-rich region or other amino acid residues contained in the aforementioned DcR3 variant, the DcR3 variant including any sugar chain is also included in the present invention.

A glycoprotein has one or two or more sugar chains. When the glycoprotein has two or more sugar chains, the number of sugar chains of the glycoprotein may be one or two or more. As the sugar chain possessed by the glycoprotein, for example, a sugar chain that N-glycosidically bonds to an amino acid residue (eg, asparagine residue, etc.) of a peptide or protein, an amino acid residue (eg, serine residue, threonine residue) of a peptide or protein and the like) a sugar chain having an O-glycosidic bond to it. Examples of the O-type sugar chain include Core 1 and Core 2, and the N-type sugar chain includes, for example, hymannose, hybrid, and complex sugar chains, but is preferably a complex sugar chain.

4. DcR3 variants comprising an Fc region

As the DcR3 variant of the present invention, a homologous or heterologous peptide, polypeptide or protein is directly on the N-terminal side or the C-terminal side of a chimeric cysteine-rich region (or a chimeric type containing another region is a cysteine-rich region) , or, if necessary, a protein bound or fused through an appropriate peptide linker is also included. The number of amino acids constituting the peptide linker is not particularly limited, and examples thereof include 4, 5, 6, or 15 amino acids.

As a polypeptide or protein to bind or fuse to a chimeric cysteine-rich region, for example, an immunoglobulin constant region or Fc region, a peptide binding to FcRn (neonatal Fc receptor), albumin, protein A, protein G, β -Galactosidase, glutathione-S-transferase (GST), maltose binding protein, polyhistidine, polypeptides or proteins such as FLAG peptide are mentioned, Preferably, the Fc region of an immunoglobulin or its variant (mutant) Fc region), more preferably a mammal-derived immunoglobulin Fc region or a variant thereof (mutant Fc region).

As the Fc region of an immunoglobulin (also referred to as an antibody), the Fc region of a human immunoglobulin is preferable for use in humans. Examples of classes and subclasses of immunoglobulins include, but are not limited to, IgG, IgD, IgE, IgM, IgA, IgG1, IgG2, IgG2a, IgG2b, IgG2c, IgG3, IgG4, or IgA1, and can be used in humans If so, it is preferable to use the class and subclass of human immunoglobulin. In addition, when an immunoglobulin Fc region is used as a polypeptide or protein to bind or fuse to a chimeric cysteine-rich region, the immunoglobulin Fc region is preferably bound or fused to the C-terminal side of the chimeric cysteine-rich region. .

Immunoglobulin is composed of heavy and light chain polypeptides, and the constant region of the heavy chain of human IgG is, in order from the N-terminus, composed of a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain. The Fc region of IgG in the present invention also includes a region in which a CH2 domain and a CH3 domain are combined, and a region in which a part or all of a hinge domain and a CH2 domain and a CH3 domain are combined. Each domain contained in the Fc region of IgG in this invention can be specified by the number of the EU index. Specifically, the hinge domain is specified by EU indexes 216th to 230th, CH2 domains 231th to 340th, and CH3 by EU indexes 341th to 447th, respectively.

In addition, to a polypeptide or protein that is bound or fused to a chimeric cysteine-rich region, the biological activity of the DcR3 variant or its properties, in vivo dynamics such as blood half-life, or physical or chemical properties such as protein stability. Also included are polypeptides or proteins modified by substitution, deletion, insertion or addition of one or more amino acids. Modifications of polypeptides or proteins that bind or fuse to a chimeric cysteine-rich region include natural mutations and artificial amino acid substitutions, deletions, insertions or additions. The modified polypeptide or protein includes, for example, a variant Fc region comprising an amino acid sequence in which one or several amino acids are deleted, substituted, inserted or added in the amino acid sequence of the Fc region of an immunoglobulin. As the amino acid sequence of the variant Fc region, for example, in the amino acid sequence of the Fc region of an immunoglobulin, one or more, preferably 2 to 30, more preferably 2 to 10, particularly preferably 2 An amino acid sequence in which 5 to 5 amino acids are substituted, deleted, inserted or added, or an amino acid sequence having 80% or more, preferably 85% or more, more preferably 90% or more identity to the amino acid sequence of the Fc region of an immunoglobulin; can be heard

The addition or deletion of the sugar chain addition site described above is also included in the modification of the polypeptide or protein to bind or fuse to the chimeric cysteine-rich region. For example, a sugar chain addition site can be added by adding or inserting a polypeptide comprising an N-type sugar chain addition site. As a specific polypeptide sequence, GGNGT or YGNGT containing 5 amino acids is mentioned [International Patent Publication No. 2014/153111].

Human IgG1 substitutions that reduce or eliminate complement-dependent cytotoxicity (CDC) activity include the EU index: Leu at position 234 (L234), Leu at position 235 (L235), Asp (D265) at position 265, Asp (D265) at position 270 Asp (D265), Lys at position 322 (K322), Pro (P329) at position 329, and Pro (P331) at position 331 of 1 or 2 or more amino acids selected from, such as substitutions in other amino acids, etc. , Specific examples include substitutions in Ala of one or more amino acids selected from L234, L235, D270, K322, P329, P331, etc., and substitutions in Ser or Gly of P331, which are indicated by the EU index [ J. Immunol., 2000, 164: p. 4178-4184, Cell. Immunol., 2000, 200: p. 16-26].

As a substitution of human IgG1 that reduces or eliminates effector activity such as antibody-dependent cytotoxicity (ADCC) activity and antibody-dependent cellular phagocytosis (ADCP), Asn (N297) at position 297, Leu at position 234 ( L234), Leu at position 235 (L235), Gly at position 237 (G237), Cys at position 226 (C226), Cys at position 229 (C229), Pro at position 238 (P238), Glu at position 233 (E233) , Ser (S267) at position 267, Leu (L328) at position 328, Pro (P331) at position 331, and the like, substitution of 1 or 2 or more amino acids with other amino acids, specifically EU index N297 at Ala (N297A), N297 at Gln (N297Q), N297 at Gly (N297G), or L234 at Ala (L234A)/L235 at Ala, represented by and amino acid substitutions such as (L235A)/G237 substitution in Ala (G237A). In addition, "/" means "and" (the same applies hereinafter).

In addition, as a modification to increase the binding activity to FcγRIIb, which is an inhibitory receptor, the substitution of Asp in Gly at position 236 (G236D), the substitution of Phe in Leu at position 328 (L328F), the 239th position represented by the EU index. substitution of Ser at Asp (S239D), and substitution of Ser at position 267 at Glu (S267E) [Curr. Opin. Cell. Biol., 2009, 20: p.685-691]. In addition, as a formula to enhance binding to FcRn (neonatal Fc receptor) in an environment with a low pH in the endosome, to avoid the loss of the antibody, and to extend the blood half-life, the 250th Thr represented by the EU index substitution in Gin (T250Q), substitution of Met at position 428 at Leu (M428L), substitution of Met at position 252 at Tyr (M252Y), substitution of Ser at position 254 at Thr (S254T), at position 256 Thr substitution in Glu (T256E), 252th Met substitution in Tyr (M252Y)/254th Ser substitution in Thr (S254T)/256th Thr substitution in Glu (T256E), 428 Substitution of Met in Leu (M428L)/434th Asn in Ser (N434S), 434th Asn substitution in Ala (N434A), 434th Asn substitution in His (N434H) et al. [J. Immunol., 2009, 182: p.7663-7671, MAbs, 2017, 9: p.844-853].

In a preferred embodiment, the mutant Fc region has Ser at position 220 indicated by the EU index in the amino acid sequence of the heavy chain of an antibody belonging to the human IgG1 subclass. As a variant Fc region related to this form, for example, from the constant region (SEQ ID NO: 153) of the heavy chain of human IgG1, the Fc region (EU index) except for the 216th Glu represented by the EU index contained in the CH1 domain and the hinge domain. Of positions 217 to 447), the Fc region of an immunoglobulin (hereinafter referred to as g1S, sequence No. 72), in the Fc region (EU index 216th to 447th) excluding the CH1 domain from the constant region (SEQ ID NO: 153) of the heavy chain of human IgG1, the amino acid sequence in which Cys at position 220 is substituted with Ser (C220S) and an immunoglobulin Fc region (hereinafter referred to as Eg1S, SEQ ID NO: 156) and the like. When the DcR3 variant of the present invention includes a mutant Fc region according to this form, as the first chimeric cysteine-rich region included in the DcR3 variant of the present invention, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG, and A first chimeric cysteine-rich region in which CRD4 of wild-type DcR3 is substituted with CRD4 of OPG is preferred. In addition, when the DcR3 variant of the present invention includes a mutant Fc region according to this form, the second chimeric cysteine-rich region contained in the DcR3 variant of the present invention is a substitution of Glu at position 57 in another amino acid; Alternatively, it is preferable to have a substitution in another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58. The other amino acid substituted with Glu at position 57 is preferably selected from Lys, Leu, Arg, Val, Ala, Phe, His, Ile and Met, and more preferably selected from Lys, Leu, Arg and Val. The other amino acid substituted with Arg at position 58 is preferably selected from Asp, Glu and Thr, and more preferably selected from Asp and Glu. In the case of combining a substitution at another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58, Lys, Leu, Arg, Val, Ala, Phe, His, Ile, or Met for Glu at position 57 It is preferable to combine the substitution of Arg at position 58 with substitution at Asp, Glu or Thr at position 57, and Substitution at Lys, Leu, Arg or Val of Glu at position 57 and Asp or Glu at position 58 It is more preferable to combine the substitution of Glu at position 57, and it is still more preferable to combine the substitution of Lys or Arg at position 57 and substitution at position 58 of Arg at Asp or Glu.

In another preferred embodiment, the mutant Fc region is, in the amino acid sequence of the heavy chain of an antibody belonging to the human IgG4 subclass, the substitution of Ser at position 228 represented by the EU index to Pro, and Leu at position 235 represented by the EU index It has a substitution with Glu and a substitution of Lys for Arg at position 409 represented by the EU index. As a variant Fc region related to this form, for example, the CH1 domain is removed from the constant region (SEQ ID NO: 154) of the heavy chain of human IgG4, and the 228th position indicated by the EU index described in International Patent Application Laid-Open No. 2006/33386 and Fc (hereinafter referred to as g4PEK, SEQ ID NO: 74) having an amino acid sequence in which Ser is substituted for Pro, Leu at position 235 is Glu, and Arg at position 409 is substituted with Lys. When the DcR3 variant of the present invention includes a mutant Fc region according to this form, as the first chimeric cysteine-rich region included in the DcR3 variant of the present invention, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG, and A first chimeric cysteine-rich region in which CRD4 of wild-type DcR3 is substituted with CRD4 of OPG is preferred. In addition, when the DcR3 variant of the present invention includes a mutant Fc region according to this form, the second chimeric cysteine-rich region contained in the DcR3 variant of the present invention is a substitution of Glu at position 57 in another amino acid; Alternatively, it is preferable to have a substitution in another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58. The other amino acid substituted with Glu at position 57 is preferably selected from Lys, Leu, Arg, Val, Ala, Phe, His, Ile and Met, and more preferably selected from Lys, Leu, Arg and Val. The other amino acid substituted with Arg at position 58 is preferably selected from Asp, Glu and Thr, and more preferably selected from Asp and Glu. In the case of combining a substitution at another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58, Lys, Leu, Arg, Val, Ala, Phe, His, Ile, or Met for Glu at position 57 It is preferable to combine the substitution of Arg at position 58 with substitution at Asp, Glu or Thr at position 57, and Substitution at Lys, Leu, Arg or Val of Glu at position 57 and Asp or Glu at position 58 It is more preferable to combine the substitution of Glu at position 57, and it is still more preferable to combine the substitution of Lys or Arg at position 57 and substitution at position 58 of Arg at Asp or Glu.

In another preferred embodiment, the variant Fc region is, in the amino acid sequence of the heavy chain of an antibody belonging to the human IgG1 subclass, the substitution of Leu at position 234 represented by the EU index with Ala and the Leu at position 235 represented by the EU index is replaced with Ala, and Gly at position 237 represented by the EU index is replaced with Ala. As a variant Fc region according to this form, for example, among the Fc regions of g1S represented by SEQ ID NO: 72 or SEQ ID NO: 156, Leu at position 234 is Ala, Leu at position 235 is Ala, and the Fc region of an immunoglobulin having an amino acid sequence in which Gly at position 237 is substituted with Ala (hereinafter referred to as g1S LALAGA or Eg1S LALAGA; SEQ ID NOs: 162 and 164) and the like. When the DcR3 variant of the present invention includes a mutant Fc region according to this form, as the first chimeric cysteine-rich region included in the DcR3 variant of the present invention, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG, and A first chimeric cysteine-rich region in which CRD4 of wild-type DcR3 is substituted with CRD4 of OPG is preferred. In addition, when the DcR3 variant of the present invention includes a mutant Fc region according to this form, the second chimeric cysteine-rich region contained in the DcR3 variant of the present invention is a substitution of Glu at position 57 in another amino acid; Alternatively, it is preferable to have a substitution in another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58. The other amino acid substituted with Glu at position 57 is preferably selected from Lys, Leu, Arg, Val, Ala, Phe, His, Ile and Met, and more preferably selected from Lys, Leu, Arg and Val. The other amino acid substituted with Arg at position 58 is preferably selected from Asp, Glu and Thr, and more preferably selected from Asp and Glu. In the case of combining a substitution at another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58, Lys, Leu, Arg, Val, Ala, Phe, His, Ile, or Met for Glu at position 57 It is preferable to combine the substitution of Arg at position 58 with substitution at Asp, Glu or Thr at position 57, and Substitution at Lys, Leu, Arg or Val of Glu at position 57 and Asp or Glu at position 58 It is more preferable to combine the substitution of Glu at position 57, and it is still more preferable to combine the substitution of Lys or Arg at position 57 and substitution at position 58 of Arg at Asp or Glu.

In another preferred embodiment, the variant Fc region has a substitution of Ala for Asn at position 434 indicated by the EU index in the amino acid sequence of the heavy chain of an antibody belonging to the human IgG1 subclass. As a mutant Fc region according to this form, for example, an immunoglobulin having an amino acid sequence in which Asn at position 434 is substituted with Ala among g1S represented by SEQ ID NO: 72 or Eg1S Fc region represented by SEQ ID NO: 156 of the Fc region (hereinafter, referred to as g1S N434A or Eg1S N434A; SEQ ID NOs: 312 and 160), g1S LALAGA shown in SEQ ID NO: 162, or Eg1S LALAGA shown in SEQ ID NO: 164 among the Fc regions of the Fc region, the 434th Asn is and the Fc region of an immunoglobulin having an amino acid sequence substituted with Ala (hereinafter referred to as g1S LALAGANA or Eg1S LALAGANA, SEQ ID NOs: 313 and 166) and the like. When the DcR3 variant of the present invention includes a mutant Fc region according to this form, as the first chimeric cysteine-rich region included in the DcR3 variant of the present invention, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG, and A first chimeric cysteine-rich region in which CRD4 of wild-type DcR3 is substituted with CRD4 of OPG is preferred. In addition, when the DcR3 variant of the present invention includes a mutant Fc region according to this form, the second chimeric cysteine-rich region contained in the DcR3 variant of the present invention is a substitution of Glu at position 57 in another amino acid; Alternatively, it is preferable to have a substitution in another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58. The other amino acid substituted with Glu at position 57 is preferably selected from Lys, Leu, Arg, Val, Ala, Phe, His, Ile and Met, and more preferably selected from Lys, Leu, Arg and Val. The other amino acid substituted with Arg at position 58 is preferably selected from Asp, Glu and Thr, and more preferably selected from Asp and Glu. In the case of combining a substitution at another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58, Lys, Leu, Arg, Val, Ala, Phe, His, Ile, or Met for Glu at position 57 It is preferable to combine the substitution of Arg at position 58 with substitution at Asp, Glu or Thr at position 57, and Substitution at Lys, Leu, Arg or Val of Glu at position 57 and Asp or Glu at position 58 It is more preferable to combine the substitution of Glu at position 57, and it is still more preferable to combine the substitution of Lys or Arg at position 57 and substitution at position 58 of Arg at Asp or Glu.

In another preferred embodiment, the variant Fc region is, in the amino acid sequence of the heavy chain of an antibody belonging to the human IgG1 subclass, Met at position 252 represented by the EU index, substituted with Tyr, and Ser at position 254 represented by the EU index of Thr, and the 256th Thr represented by the EU index is substituted with Glu. As a variant Fc region according to this form, for example, among the Fc regions of g1S represented by SEQ ID NO: 72 or SEQ ID NO: 156, Met at position 252 is Tyr, Ser at position 254 is Thr, and the Fc region of an immunoglobulin having an amino acid sequence in which Thr at position 256 is substituted with Glu (hereinafter referred to as g1S YTE or Eg1S YTE; SEQ ID NOs: 311 and 158) and the like. When the DcR3 variant of the present invention includes a mutant Fc region according to this form, as the first chimeric cysteine-rich region included in the DcR3 variant of the present invention, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG, and A first chimeric cysteine-rich region in which CRD4 of wild-type DcR3 is substituted with CRD4 of OPG is preferred. In addition, when the DcR3 variant of the present invention includes a mutant Fc region according to this form, the second chimeric cysteine-rich region contained in the DcR3 variant of the present invention is a substitution of Glu at position 57 in another amino acid; Alternatively, it is preferable to have a substitution in another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58. The other amino acid substituted with Glu at position 57 is preferably selected from Lys, Leu, Arg, Val, Ala, Phe, His, Ile and Met, and more preferably selected from Lys, Leu, Arg and Val. The other amino acid substituted with Arg at position 58 is preferably selected from Asp, Glu and Thr, and more preferably selected from Asp and Glu. In the case of combining a substitution at another amino acid for Glu at position 57 and a substitution at another amino acid for Arg at position 58, Lys, Leu, Arg, Val, Ala, Phe, His, Ile, or Met for Glu at position 57 It is preferable to combine the substitution of Arg at position 58 with substitution at Asp, Glu or Thr at position 57, and Substitution at Lys, Leu, Arg or Val of Glu at position 57 and Asp or Glu at position 58 It is more preferable to combine the substitution of Glu at position 57, and it is still more preferable to combine the substitution of Lys or Arg at position 57 and substitution at position 58 of Arg at Asp or Glu.

The variant Fc region includes, but is not limited to, the amino acid sequence set forth in, for example, SEQ ID NOs: 72, 74, 156, 158, 160, 162, 164, 166, 311, 312 or 313, or the corresponding amino acids and a variant Fc region comprising a sequence.

The peptide linker added to the N-terminal or C-terminal side of the chimeric cysteine-rich region to further link the same or heterogeneous peptide, polypeptide or protein is not limited, but for example, an IEGRMD linker or a GS linker. A peptide linker, a chemical linker, etc. are mentioned.

As the DcR3 variant of the present invention, most preferably, a DcR3 variant comprising a first or second chimeric cysteine-rich region and an Fc region or a variant Fc region may be mentioned.

In one embodiment, as the amino acid sequence contained in the DcR3 variant of the present invention, for example, the amino acid sequence shown in SEQ ID NOs: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44 or 46, or and amino acid sequences in which 1 to 30 amino acids are deleted, substituted, inserted or added in the amino acid sequence. In this aspect, the DcR3 variant of the present invention may consist of the amino acid sequence or may include the amino acid sequence, but preferably consist of the amino acid sequence. In addition, the amino acid sequence includes HBD.

In another embodiment, the amino acid sequence contained in the DcR3 variant of the present invention is, for example, SEQ ID NO: 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 180, 182, 184 , 186, 188, 270, 272, 274, 276, 278, 280, 282, 284 or 286, or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence can be heard In this aspect, the DcR3 variant of the present invention may consist of the amino acid sequence or may include the amino acid sequence, but preferably consist of the amino acid sequence. In addition, the amino acid sequence does not include HBD.

In another embodiment, the amino acid sequence contained in the DcR3 variant of the present invention is, for example, SEQ ID NO: 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 180, 182, 184 , 186, 188, 270, 272, 274, 276, 278, 280, 282, 284 or 286, or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence and an amino acid sequence comprising the amino acid sequence set forth in SEQ ID NOs: 72, 74, 156, 158, 160, 162, 164, 166, 311, 312 or 313 as the variant Fc region.

In another embodiment, as the amino acid sequence contained in the DcR3 variant of the present invention, for example, SEQ ID NOs: 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 150, 168, 170, 172, 174, 176, 178, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 288, 290, 292, 294, 296, 298, 300, 302, 304, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, The amino acid sequence described in 334, 335, 336 or 337, or an amino acid sequence in which 1 to 30 amino acids are deleted, substituted, inserted or added in the amino acid sequence. In this aspect, the DcR3 variant of the present invention may consist of the amino acid sequence or may include the amino acid sequence, but preferably consist of the amino acid sequence. In addition, the amino acid sequence includes an Fc region or a variant thereof (variant Fc region).

SEQ ID NO: 76 is the amino acid sequence of chimeric B-Fc (IEGRMD g1S) (DcR3 variant in which chimeric B, IEGRMD linker, and Fc (g1S) are fused), SEQ ID NO: 78 is chimeric C-Fc (IEGRMD) The amino acid sequence of g1S) (chimeric C, IEGRMD linker, and DcR3 variant fused with Fc (g1S)), SEQ ID NO: 80, is chimeric A-Fc (IEGRMD g1S) (chimeric A, IEGRMD linker, and Fc ( The amino acid sequence of DcR3 variant fused with g1S), SEQ ID NO: 82 is the amino acid sequence of chimeric A-Fc (g4PEK) (DcR3 variant fused with chimera A and Fc (g4PEK)), SEQ ID NO: 84, The amino acid sequence of 103-123OPG-Fc (g4PEK) (103-123OPG and Fc (g4PEK) fused DcR3 variant), SEQ ID NO: 86 is N131S / N144S-Fc (g4PEK) (N131S / N144S and Fc ( The amino acid sequence of DcR3 variant fused with g4PEK), SEQ ID NO: 88, is the amino acid sequence of T133A / S146A-Fc (g4PEK) (DcR3 variant fused with T133A / S146A and Fc (g4PEK)), SEQ ID NO: 90 Silver, the amino acid sequence of N131S / N144S / N157S-Fc (g4PEK) (N131S / N144S / N157S and Fc (g4PEK) fused DcR3 variant), SEQ ID NO: 92 is T133A / S146A / T159A-Fc (g4PEK) The amino acid sequence of (T133A/S146A/T159A and DcR3 variant in which Fc (g4PEK) is fused), SEQ ID NO: 94, is chimeric A-E57K-Fc (g4PEK) (chimeric A-E57K and Fc (g4PEK) fused) The amino acid sequence of the DcR3 variant), SEQ ID NO: 96, is the amino acid sequence of the chimeric A-E57L-Fc (g4PEK) (the DcR3 variant in which the chimeric A-E57L and Fc (g4PEK) are fused), SEQ ID NO: 98, Chimeric A-R60K-Fc (g4PEK) (chimeric A-R60K and Fc (g4PEK) fused DcR3 modification body) amino acid sequence, SEQ ID NO: 150 is the amino acid sequence of chimeric A-Fc(g1S) (DcR3 variant in which chimera A and Fc(g1S) are fused), SEQ ID NO: 168 is chimeric A-Fc(Eg1S) The amino acid sequence of (chimera A and DcR3 variant fused with Fc (Eg1S)), SEQ ID NO: 170, is a DcR3 variant fused with chimera A-Fc (Eg1S-YTE) (chimera A and Fc (Eg1S YTE)) ), the amino acid sequence of SEQ ID NO: 172 is the amino acid sequence of chimeric A-Fc (Eg1S-N434A) (chimeric A and Fc (Eg1S-N434A) fused DcR3 variant), SEQ ID NO: 174 is chimeric A-Fc The amino acid sequence, SEQ ID NO: 176 of (g1S-LALAGA) (chimera A and Fc (g1S-LALAGA) fused DcR3 variant) is chimeric A-Fc (Eg1S-LALAGA) (chimera A and Fc (Eg1S-) The amino acid sequence, SEQ ID NO: 178 of the DcR3 variant fused to LALAGA) is the amino acid sequence and sequence of the chimeric A-Fc (Eg1S-LALAGANA) (chimera A and Fc (Eg1S-LALAGANA) fused DcR3 variant) No. 190 is the amino acid sequence of chimeric A-E57R-Fc(g4PEK) (DcR3 variant in which chimeric A-E57R and Fc(g4PEK) are fused), SEQ ID NO: 192 is chimeric A-E57V-Fc(g4PEK) ( The amino acid sequence of chimeric A-E57V and Fc (g4PEK) fused DcR3 variant), SEQ ID NO: 194, is chimeric A-E57K_R58D-Fc (g4PEK) (chimeric A-E57K_R58D with Fc (g4PEK) fused to DcR3 The amino acid sequence of the variant), SEQ ID NO: 196, is the amino acid sequence of chimeric A-E57K_R58E-Fc(g4PEK) (chimeric A-E57K_R58E and Fc (g4PEK) fused DcR3 variant), SEQ ID NO: 198 is chimeric A -E57R_R58D-Fc (g4PEK) (chimeric A-E57R_R58D and Fc (g4PEK) fused DcR3 variant) amino acid The sequence, SEQ ID NO: 200 is the amino acid sequence of chimeric A-E57K-Fc (Eg1S) (DcR3 variant obtained by fusion of chimeric A-E57K and Fc (Eg1S)), SEQ ID NO: 202 is chimeric A-E57L-Fc ( The amino acid sequence, SEQ ID NO: 204 of Eg1S) (chimeric A-E57L and DcR3 variant fused with Fc (Eg1S)) is chimeric A-E57R-Fc (Eg1S) (chimeric A-E57R and Fc (Eg1S) The amino acid sequence of the fused DcR3 variant), SEQ ID NO: 206, is the amino acid sequence, SEQ ID NO: 208 of the chimeric A-E57V-Fc (Eg1S) (the DcR3 variant fused with the chimeric A-E57V and Fc (Eg1S)) , the amino acid sequence of chimeric A-E57K_R58D-Fc(Eg1S) (chimeric A-E57K_R58D and Fc(Eg1S) fused DcR3 variant), SEQ ID NO: 210 is chimeric A-E57K_R58E-Fc(Eg1S) (chimeric A- The amino acid sequence of E57K_R58E and Fc (Eg1S) fused DcR3 variant), SEQ ID NO: 212, is chimeric A-E57R_R58D-Fc (Eg1S) (chimeric A-E57R_R58D and Fc (Eg1S) fused DcR3 variant) The amino acid sequence of SEQ ID NO: 214 is the amino acid sequence of chimeric A-E57K-Fc(Eg1S YTE) (chimeric A-E57K and Fc (Eg1S YTE) fused DcR3 variant), SEQ ID NO: 216 is chimeric A- The amino acid sequence of E57L-Fc (Eg1S YTE) (DcR3 variant obtained by fusion of chimeric A-E57L and Fc (Eg1S YTE)), SEQ ID NO: 218, is chimeric A-E57R-Fc (Eg1S YTE) (chimeric A-E57R) And, the amino acid sequence of DcR3 variant fused with Fc (Eg1S YTE)), SEQ ID NO: 220, chimeric A-E57V-Fc (Eg1S YTE) (chimeric A-E57V and Fc (Eg1S YTE) fused DcR3 dog The amino acid sequence of the variant), SEQ ID NO: 222, is chimeric A-E57K_R58D-Fc(Eg1S YTE) (chimeric A-E57K_R) The amino acid sequence of 58D and Fc (Eg1S YTE) fused DcR3 variant), SEQ ID NO: 224, is chimeric A-E57K_R58E-Fc (Eg1S YTE) (chimeric A-E57K_R58E and Fc (Eg1S YTE) fused to DcR3 The amino acid sequence of the variant), SEQ ID NO: 226, is the amino acid sequence of chimeric A-E57R_R58D-Fc(Eg1S YTE) (chimeric A-E57R_R58D and Fc (Eg1S YTE) fused DcR3 variant), SEQ ID NO: 228, The amino acid sequence of chimeric A-E57K-Fc(Eg1S N434A) (DcR3 variant obtained by fusing chimeric A-E57K and Fc (Eg1S N434A)), SEQ ID NO: 230, is chimeric A-E57L-Fc(Eg1S N434A) (chimera) The amino acid sequence of A-E57L and Fc (Eg1S N434A) fused DcR3 variant), SEQ ID NO: 232, is the chimeric A-E57R-Fc (Eg1S N434A) (chimeric A-E57R and Fc (Eg1S N434A) fusion) The amino acid sequence of the DcR3 variant), SEQ ID NO: 234, is the amino acid sequence of the chimeric A-E57V-Fc (Eg1S N434A) (the DcR3 variant in which the chimeric A-E57V and Fc (Eg1S N434A) are fused), SEQ ID NO: 236 Silver, the amino acid sequence of chimeric A-E57K_R58D-Fc(Eg1S N434A) (DcR3 variant obtained by fusion of chimeric A-E57K_R58D and Fc (Eg1S N434A)), SEQ ID NO: 238 is chimeric A-E57K_R58E-Fc(Eg1S N434A) The amino acid sequence of (chimeric A-E57K_R58E and DcR3 variant fused with Fc (Eg1S N434A)), SEQ ID NO: 240, is chimeric A-E57R_R58D-Fc (Eg1S N434A) (chimeric A-E57R_R58D and Fc (Eg1S N434A) The amino acid sequence of the DcR3 variant fused to), SEQ ID NO: 242, is the chimeric A-E57K-Fc (Eg1S LALAGA) (the DcR3 variant fused with the chimeric A-E57K and Fc (Eg1S LALAGA)) The amino acid sequence, SEQ ID NO: 244 is the amino acid sequence of chimeric A-E57L-Fc (Eg1S LALAGA) (DcR3 variant obtained by fusion of chimeric A-E57L and Fc (Eg1S LALAGA)), SEQ ID NO: 246 is chimeric A-E57R -Fc (Eg1S LALAGA) (chimeric A-E57R and Fc (Eg1S LALAGA) fused DcR3 variant) amino acid sequence, SEQ ID NO: 248, chimeric A-E57V-Fc (Eg1S LALAGA) (chimeric A-E57V and , The amino acid sequence of DcR3 variant fused with Fc (Eg1S LALAGA)), SEQ ID NO: 250 is chimeric A-E57K_R58D-Fc (Eg1S LALAGA) (chimeric A-E57K_R58D and DcR3 variant fused with Fc (Eg1S LALAGA) ), the amino acid sequence of SEQ ID NO: 252 is the amino acid sequence of chimeric A-E57K_R58E-Fc (Eg1S LALAGA) (chimeric A-E57K_R58E and Fc (Eg1S LALAGA) fused DcR3 variant), SEQ ID NO: 254 is chimeric A -E57R_R58D-Fc (Eg1S LALAGA) (chimeric A-E57R_R58D and Fc (Eg1S LALAGA) fused DcR3 variant) amino acid sequence, SEQ ID NO: 256 is chimeric A-E57K-Fc (Eg1S LALAGANA) (chimeric A- The amino acid sequence of E57K and Fc (Eg1S LALAGANA) fused DcR3 variant), SEQ ID NO: 258, is chimeric A-E57L-Fc (Eg1S LALAGANA) (chimeric A-E57L and DcR3 fused with Fc (Eg1S LALAGANA) The amino acid sequence of the variant), SEQ ID NO: 260, is the amino acid sequence of chimeric A-E57R-Fc (Eg1S LALAGANA) (chimeric A-E57R and Fc (Eg1S LALAGANA) fused DcR3 variant), SEQ ID NO: 262 is, Amino acid sequence, sequence number of chimeric A-E57V-Fc (Eg1S LALAGANA) (DcR3 variant obtained by fusion of chimeric A-E57V and Fc (Eg1S LALAGANA)) No. 264 is the amino acid sequence of chimeric A-E57K_R58D-Fc(Eg1S LALAGANA) (DcR3 variant obtained by fusing chimeric A-E57K_R58D and Fc (Eg1S LALAGANA)), SEQ ID NO: 266 is the chimeric A-E57K_R58E-Fc(Eg1S) The amino acid sequence of LALAGANA) (chimeric A-E57K_R58E and DcR3 variant fused with Fc (Eg1S LALAGANA)), SEQ ID NO: 268, is chimeric A-E57R_R58D-Fc (Eg1S LALAGANA) (chimeric A-E57R_R58D and Fc (Eg1S) The amino acid sequence, SEQ ID NO: 288 of the DcR3 variant fused with LALAGANA) is the amino acid sequence and sequence of the chimeric A-E57A-Fc (g4PEK) (the DcR3 variant fused with the chimeric A-E57A and Fc (g4PEK)) No. 290 is the amino acid sequence of chimeric A-E57F-Fc (g4PEK) (DcR3 variant obtained by fusion of chimeric A-E57F and Fc (g4PEK)), SEQ ID NO: 292 is chimeric A-E57H-Fc (g4PEK) ( The amino acid sequence of chimeric A-E57H and Fc (g4PEK) fused DcR3 variant), SEQ ID NO: 294, is chimeric A-E57I-Fc (g4PEK) (chimeric A-E57I and Fc (g4PEK) fused DcR3 The amino acid sequence of the variant), SEQ ID NO: 296, is the amino acid sequence of chimeric A-E57M-Fc (g4PEK) (DcR3 variant obtained by fusion of chimeric A-E57M and Fc (g4PEK)), SEQ ID NO: 298 is chimeric A -E57K_R58T-Fc(g4PEK) (chimeric A-E57K_R58T and DcR3 variant fused with Fc (g4PEK)) amino acid sequence, SEQ ID NO: 300 is chimeric A-E57L_R58E-Fc(g4PEK) (chimeric A-E57L_R58E, The amino acid sequence of DcR3 variant fused with Fc (g4PEK)), SEQ ID NO: 302, is the amino acid of chimeric A-E57V_R58T-Fc (g4PEK) (chimeric A-E57V_R58T and DcR3 variant fused with Fc (g4PEK)) The acid sequence, SEQ ID NO: 304 is the amino acid sequence of chimeric A-E57V_R58E-Fc(g4PEK) (DcR3 variant obtained by fusing chimeric A-E57V_R58E and Fc (g4PEK)), SEQ ID NO: 314 is chimeric A-Fc (g1S) The amino acid sequence of YTE) (chimera A and Fc (g1S YTE) fused DcR3 variant), SEQ ID NO: 315, is chimeric A-Fc (g1S N434A) (chimera A and Fc (g1S N434A) fused DcR3 The amino acid sequence of the variant), SEQ ID NO: 316, is the amino acid sequence of chimeric A-Fc (g1S LALAGANA) (chimeric A and Fc (g1S LALAGANA) fused DcR3 variant), SEQ ID NO: 317 is chimeric A-E57K -Fc (g1S YTE) (chimeric A-E57K and Fc (g1S YTE) fused DcR3 variant) amino acid sequence, SEQ ID NO: 318, chimeric A-E57L-Fc (g1S YTE) (chimeric A-E57L and , The amino acid sequence of DcR3 variant fused with Fc (g1S YTE)), SEQ ID NO: 319, is chimeric A-E57R-Fc (g1S YTE) (chimeric A-E57R and DcR3 variant fused with Fc (g1S YTE)) ), the amino acid sequence of SEQ ID NO: 320 is the amino acid sequence of chimeric A-E57V-Fc (g1S YTE) (chimeric A-E57V and Fc (g1S YTE) fused DcR3 variant), SEQ ID NO: 321 is chimeric A The amino acid sequence of -E57K_R58D-Fc(g1S YTE) (DcR3 variant obtained by fusion of chimeric A-E57K_R58D and Fc(g1S YTE)), SEQ ID NO: 322 is chimeric A-E57K_R58E-Fc(g1S YTE) (chimeric A- The amino acid sequence of E57K_R58E and DcR3 variant fused with Fc (g1S YTE)), SEQ ID NO: 323, is chimeric A-E57R_R58D-Fc (g1S YTE) (chimeric A-E57R_R58D with Fc (g1S YTE) fused to DcR3 The amino acid sequence of the variant), SEQ ID NO: 324, is chimeric A-E57K-Fc (g1 The amino acid sequence of S N434A) (chimeric A-E57K and Fc (g1S N434A) fused DcR3 variant), SEQ ID NO: 325, is chimeric A-E57L-Fc (g1S N434A) (chimeric A-E57L and Fc ( The amino acid sequence of the DcR3 variant fused with g1S N434A), SEQ ID NO: 326 is the amino acid of the chimeric A-E57R-Fc (g1S N434A) (the DcR3 variant fused with the chimeric A-E57R and Fc (g1S N434A)) The sequence, SEQ ID NO: 327 is the amino acid sequence of chimeric A-E57V-Fc (g1S N434A) (DcR3 variant obtained by fusion of chimeric A-E57V and Fc (g1S N434A)), SEQ ID NO: 328 is chimeric A-E57K_R58D- The amino acid sequence of Fc (g1S N434A) (DcR3 variant fused with chimeric A-E57K_R58D and Fc (g1S N434A)), SEQ ID NO: 329, is chimeric A-E57K_R58E-Fc (g1S N434A) (chimeric A-E57K_R58E and The amino acid sequence of DcR3 variant fused with Fc (g1S N434A)), SEQ ID NO: 330, is chimeric A-E57R_R58D-Fc (g1S N434A) (chimeric A-E57R_R58D and DcR3 variant fused with Fc (g1S N434A)) The amino acid sequence of SEQ ID NO: 331 is the amino acid sequence of chimeric A-E57K-Fc (g1S LALAGANA) (DcR3 variant obtained by fusing chimeric A-E57K and Fc (g1S LALAGANA)), SEQ ID NO: 332 is chimeric A- The amino acid sequence, SEQ ID NO: 333 of E57L-Fc (g1S LALAGANA) (chimeric A-E57L and Fc (g1S LALAGANA) fused DcR3 variant) is chimeric A-E57R-Fc (g1S LALAGANA) (chimeric A-E57R) And, the amino acid sequence of DcR3 variant fused with Fc (g1S LALAGANA)), SEQ ID NO: 334 is chimeric A-E57V-Fc (g1S LALAGANA) (chimeric A-E57V and DcR3 fused with Fc (g1S LALAGANA) The amino acid sequence of the variant), SEQ ID NO: 335, is the amino acid sequence of the chimeric A-E57K_R58D-Fc (g1S LALA GANA) (chimeric A-E57K_R58D and Fc (g1S LALAGANA) fused DcR3 variant), SEQ ID NO: 336 is , the amino acid sequence of chimeric A-E57K_R58E-Fc (g1S LALAGANA) (DcR3 variant obtained by fusing chimeric A-E57K_R58E and Fc (g1S LALAGANA)), SEQ ID NO: 337 is the chimeric A-E57R_R58D-Fc (g1S LALAGANA) ( The amino acid sequence of the chimeric A-E57R_R58D and Fc (g1S LALAGANA) fused DcR3 variant) is shown.

In any case of the above embodiment, the variation (modification) applied to the amino acid sequence includes both natural mutations and artificial amino acid substitutions, deletions, insertions or additions. Further, in any case of the above embodiment, in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added as a sequence, 1 or 2 or more, preferably 2 to 30, more preferably is an amino acid sequence in which 2 to 10, particularly preferably 2 to 5 amino acids are substituted, deleted, inserted, or added, or 80% or more, preferably 85% or more, more preferably 90% or more with the corresponding amino acid sequence. , for example, an amino acid sequence having 93% or more, 95% or more, 97% or more, 98% or more, or 99% or more identity.

The DcR3 variant of the present invention may be further chemically modified for the purpose of changing biological activity or properties thereof, in vivo dynamics such as blood half-life, or physical or chemical properties such as protein stability.

Examples of the chemical modification include polyethylene glycol (PEG) formation, acetylation, amidation, or phosphorylation, and particularly preferably PEGylation. PEGylation is, for example, binding one or more PEG molecules to an amino acid residue having a functional group such as an N-terminal amino group of a protein, an ε amino group of Lys, a carboxyl group, a thiol group, or a hydroxyl group in the side chain.

The average molecular weight of the PEG molecule is not limited to the following, but can be used in the range of about 3000 to about 50000.

As a method of linking a PEG molecule to a DcR3 modified product, an active group such as a carboxyl group, formyl (aldehyde) group, N-hydroxysuccinimide ester group, amino group, thiol group or maleimide group at the PEG terminal portion and a method of reacting with a functional group such as an amino group, a carboxyl group, a thiol group, or a hydroxyl group having in the side chain of the DcR3 modified product.

The signal peptide is, but not limited to, when the CRD domain on the N-terminal side contained in the DcR3 variant is derived from human DcR3, the signal peptide of human DcR3 containing the amino acid sequence 1 to 29 of SEQ ID NO: 2 is mentioned. can Moreover, when the CRD domain on the N-terminal side contained in the DcR3 variant is derived from human OPG, a signal peptide derived from human OPG containing the amino acid sequence of positions 1 to 21 of SEQ ID NO: 14 is exemplified. In addition, artificial sequences, sequences derived from expression vectors, and sequences derived from other proteins suitable for host cells expressing the DcR3 variant are all included in the signal peptide of the present invention. The DcR3 variant including the signal peptide is an immature polypeptide, and in one embodiment, the signal peptide is cleaved during maturation. The DcR3 variants of the present invention include those having a different N-terminal cleavage at a position different from the predicted site for cleavage of the signal peptide.

5. Method for producing a DcR3 variant

The DcR3 variant of the present invention uses the method described in Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory Press (2001), etc., for example, by the following method, which encodes the DcR3 variant It can be prepared by expressing DNA in a host cell.

The signal peptide is, but not limited to, when the CRD domain on the N-terminal side contained in the DcR3 variant is derived from human DcR3, the signal peptide of human DcR3 containing the amino acid sequence 1 to 29 of SEQ ID NO: 2 is mentioned. can Moreover, when the CRD domain on the N-terminal side contained in the DcR3 variant is derived from human OPG, a signal peptide derived from human OPG containing the amino acid sequence of positions 1 to 21 of SEQ ID NO: 14 is exemplified. In addition, an artificial sequence, a sequence derived from an expression vector, and a sequence derived from another protein suitable for a host cell expressing the DcR3 variant can all be used in the preparation of the DcR3 variant of the present invention. The DcR3 variants of the present invention include those having a different N-terminal cleavage at a position different from the predicted site for cleavage of the signal peptide.

The DcR3 variant of the present invention may be obtained by artificially designing based on the amino acid sequence of the cysteine-rich region of the wild-type DcR3 before amino acid substitution, the amino acid sequence of the cysteine-rich region of the DcR3 variant, or the amino acid sequence of the DcR3 variant, It can also be obtained by analyzing the mutant. As the mutagenesis method, site-directed mutagenesis using a PCR method using primers is preferable (Kunkel et al., Proc. Natl. Acad. Sci. USA, 1985, 82: 488-492). In addition, a method of synthesizing the entire mutated gene, or using a primer containing the mutation, dividing the part before and after the mutation site and performing PCR, the two fragments of the region containing the mutation A method of ligating by overlap and inserting into a vector by an in-fusion cloning method (Clontech) or the like is exemplified.

As a method for identifying mutations having the desired binding modality, a method of obtaining a mutant having the target binding modality by preparing a library into which the mutation is introduced at random, displaying it in phage or yeast, etc., and screening by binding activity is mentioned. there is. Alternatively, there is a method in which a vector into which a DNA mutation for substituting a specific amino acid is introduced with another amino acid is expressed in a host cell and produced, and a mutant having the desired binding pattern is obtained. DNA encoding the DcR3 variant of the present invention can be synthesized by a DNA synthesizer by designing a nucleotide sequence encoding it from the amino acid sequence of the DcR3 variant of the present invention. It can also be isolated by PCR using human cDNA as a template.

A recombinant vector is prepared by inserting the DNA encoding the DcR3 variant of the present invention obtained above into a promoter downstream of an appropriate expression vector, and the recombinant vector is introduced into a host cell suitable for the expression vector.

As the nucleotide sequence of the DNA encoding the DcR3 variant, the base can be substituted so that the codon is optimal for expression in the host, thereby improving the production rate of the desired DcR3 variant. When preparing the DNA encoding the above-mentioned DcR3 variant, DNA encoding the signal peptide of the secreted protein is added to its 5' end to prepare, and using the DNA, a recombinant vector is prepared in the same manner as above, and the host By introducing into cells, the peptide can be secreted into the medium and produced. Examples of the signal peptide include sequences derived from DcR3 or OPG, artificial sequences, sequences derived from expression vectors, and sequences derived from other proteins suitable for host cells.

In one embodiment, as the nucleotide sequence of the DNA encoding the amino acid sequence of the DcR3 variant, for example, the nucleotide sequence shown in SEQ ID NOs: 25, 27, 29, 31, 33, 35, 37, 39, 41, 43 and 45 can be heard

In another embodiment, as the nucleotide sequence of the DNA encoding the amino acid sequence of the DcR3 variant, for example, SEQ ID NOs: 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 179, 181 , 183, 185, 187, 269, 271, 273, 275, 277, 279, 281, 283 and 285.

In another embodiment, as the base sequence of the DNA encoding the amino acid sequence of the DcR3 variant, for example, SEQ ID NOs: 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 149, 167, 169, 171, 173, 175, 177, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 287, 289, 291, The nucleotide sequences described in 293, 295, 297, 299, 301 and 303 are mentioned.

As the expression vector, any expression vector can be used as long as it contains an appropriate promoter at a position capable of autonomous replication in the host cell to be used or introduction into the chromosome and capable of transcribed DNA encoding the polypeptide.

As the host cell, any one capable of expressing a gene encoding a variant DcR3, such as yeast, insect cell, or animal cell, can be used. Preferably, yeast, animal cells, or insect cells capable of glycosylating a protein are mentioned.

As the yeast, for example, Saccharomyces genus, Schizosaccharomyces genus, Kluyveromyces genus, Trichosporon genus, Schwanniomyces genus, Pichia ( Pichia ) Genus, Candida genus, etc. microorganisms belonging to, for example, Saccharomyces cerevisiae , Schizosaccharomyces pombe ), Kluyveromyces lactis ( Kluyveromyces lactis ), Tricos Poron pullulans ( Trichosporon pullulans ), Schwanniomyces alluvius ( Schwanniomyces alluvius ), or Candida utilis ( Candida utilis ), and the like.

As insect cells, for example, Sf9, Sf21, which are ovarian cells of Spodoptera frugiperda [Baculovirus Expression Vectors, A Laboratory Manual, W. H. Freeman and Company, New York (1992)], Tricofluciani (Trichoplusia ni) ovarian cells High 5 (manufactured by Invitrogen), or Drosophila melanogaster (Drosophila melanogaster) terminal embryo-derived S2 (Schneider 2) cells (Thermo Scientific), etc. are mentioned.

As an animal cell, For example, Namalwa cells which are human cells, COS cells which are monkey cells, and CHO cells which are Chinese hamster cells [Journal of Experimental Medicine, 108, 945 (1958); Proc. Natl. Acad. Sci. USA, 60, 1275 (1968); Genetics, 55, 513 (1968); Chromosoma, 41, 129 (1973); Methods in Cell Science, 18, 115 (1996); Radiation Research, 148, 260 (1997); Proc. Natl. Acad. Sci. USA, 77, 4216 (1980); Proc. Natl. Acad. Sci. USA, 60, 1275 (1968); Cell, 6, 121 (1975); Molecular Cellgenetics, Appendix I, II (pp.883-900), CHO/DG44, CHO-K1 (ATCC No.: CCL-61), Freestyle CHO-S cells, CHO cells deficient in the dihydrofolate reductase gene [Proc . Natl. Acad. Sci. USA, 77, 4216 (1980)], CHO cells deficient in 6-fucose transferase gene (International Publication No. 2005/035586, International Publication No. 02/31140), human 293 cells (ATCC No.: CRL-1573), Freestyle 293F cells, Expi293 cells (Thermo Scientific), DUkXB11 (ATCC number: CCL-9096), Pro-5 (ATCC number: CCL-1781), CHO-S (Life Technologies, Cat#11619) , Pro-3, rat myeloma cells YB2/3HL.P2.G11.16Ag.20 (also called YB2/0), mouse myeloma cells NSO, mouse myeloma cells SP2/0-Ag14, or Syrian hamster cells. BHK, HBT5637 (Unexamined-Japanese-Patent No. 63-299), etc. are mentioned.

When yeast is used as a host cell, YEP13 (ATCC number: 37115), YEp24 (ATCC37051), YCp50 (ATCC37419), pHS19, pHS15 etc. are mentioned as an expression vector, for example.

As the promoter, any promoter that can be expressed in the yeast strain may be used. For example, a gene promoter of the corresponding system such as hexose kinase, PHO5 promoter, PGK promoter, GAP promoter, ADH promoter, gal 1 promoter, and gal 10 promoter. , a heat shock polypeptide promoter, an MFα1 promoter, or a CUP1 promoter.

Any method of introducing DNA into yeast can be used as a method for introducing a recombinant vector, for example, the electroporation method [Methods Enzymol., 194, 182 (1990)], the spheroplast method [Proc. Natl. Acad. Sci. USA, 75, 1929 (1978)], lithium acetate method [J. Bacteriology, 153, 163 (1983)], or Proc. Natl. Acad. Sci. USA, 75, the method described in 1929 (1978), etc. are mentioned.

When insect cells are used as hosts, for example, Current Protocols In Molecular Biology, Baculovirus Expression Vectors, A Laboratory Manual, W. H. Freeman and Company, New York (1992), Bio/Technology, 6 , 47 (1988), etc., can express the peptide.

That is, the recombinant virus can be obtained from the insect cell culture supernatant by co-introducing the recombinant transgenic vector and the defective baculovirus genome into insect cells, and then further infecting the insect cells with the recombinant virus to express the peptide.

As a transgenic vector used in this method, pVL1392, pVL1393 (made by Becton Dickinson), pBlueBac4.5 (made by Invitrogen) etc. are mentioned, for example.

As the baculovirus, for example, Autographa californica nuclear polyhedrosis virus (Autographa californica nuclear polyhedrosis virus) which is a virus that infects chestnut moths and insects can be used.

As a method for co-introduction of the recombinant gene transfer vector and the baculovirus into insect cells for preparing a recombinant virus, for example, a calcium phosphate method (Japanese Patent Application Laid-Open No. 2-227075) or a lipofection method [Proc] . Natl. Acad. Sci. USA, 84, 7413 (1987)] and the like.

When using as a host the S2 (Schneider 2) cell (Thermo Scientific) derived from the terminal embryo of insect cell Drosophila melanogaster, for example, Mol. Biotechnol., 2015, 10: p.914-922, etc., by introducing a gene transfer vector such as pMTBiPV5-HisA (Thermo Scientific) into host cells by the calcium phosphate method, peptides can be expressed .

When using an animal cell as a host, as an expression vector, for example, pCI mammalian expression vector (Promega), pcDNA3.1(+) (Invitrogen), pcDNA I/Amp, pcDNA I, pcDM8 (after Nakoshi Corporation), pAGE107 [Japanese Patent Laid-Open No. 3-22979, Cytotechnology, 3, 133 (1990)], pAS3-3 (Japanese Patent Laid-Open No. 2-227075), pCDM8 [Nature, 329, 840 (1987). )], pREP4 (manufactured by Invitrogen), pAGE103 [J. Biochem., 101, 1307 (1987)], pAGE210, pME18SFL3, or pKANTEX93 (International Publication No. 97/10354).

As the promoter, any one that functions in animal cells can be used. For example, the promoter of IE (immediate early) gene of cytomegalovirus (CMV), early promoter of SV40, promoter of retrovirus, metallothione phosphorus promoter, heat shock promoter, or SRα promoter; In addition, the enhancer of the IE gene of human CMV may be used together with a promoter.

As a method for introducing a recombinant vector into animal cells, any method can be used as long as it is a method for introducing DNA into animal cells. For example, the electroporation method [Cytotechnology, 3, 133 (1990)], the calcium phosphate method (Japanese Patent Laid-Open) Hei 2-227075 publication), lipofection method [Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)], or the method described in Virology, 52, 456 (1973).

Transient expression of the DcR3 variant can be performed using the expression vector of the DcR3 variant of the present invention obtained by the above method, or an expression vector obtained by modifying them.

As the host cell into which the expression vector is introduced, any host cell capable of expressing the DcR3 variant can be used, for example, COS-7 cells (ATCC No.: CRL1651) are used [Methods in Nucleic Acids Res. , CRC Press, 283 (1991)]. The introduction of the expression vector into COS-7 cells includes the DEAE-dextran method [Methods in Nucleic Acids Res., CRC Press, (1991)], or the lipofection method [Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)] and the like are used.

When using CHO-S cells or Expi293 cells (Thermo Scientific), the lipofection method [Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)] and the like are used.

A transformant that stably expresses the DcR3 variant can be obtained by using the expression vector of the DcR3 variant of the present invention obtained by the above method, or an expression vector obtained by modifying them.

After introduction of the expression vector, a transformant that stably expresses the recombinant antibody is selected by culturing in an animal cell culture medium containing a drug such as G418 sulfate (Japanese Patent Application Laid-Open No. 2-257891).

When the transformant is a transformant obtained by using a eukaryote such as yeast as a host, as a medium for culturing the transformant, the transformant contains a magnetizable carbon source, nitrogen source and/or inorganic salt, etc. As long as it is a medium capable of efficiently culturing the transformant, either a natural medium or a synthetic medium may be used.

As the carbon source, the transformant should be capable of being magnetized, for example, glucose, fructose, sucrose, molasses containing these, carbohydrates such as starch or starch hydrolyzate, organic acids such as acetic acid and propionic acid, ethanol or propanol Alcohols, such as these, etc. can be used.

Examples of the nitrogen source include ammonium salts of inorganic or organic acids such as ammonia, ammonium chloride, ammonium sulfate, ammonium acetate, and/or ammonium phosphate, other nitrogen-containing compounds, and peptone, meat extract, yeast extract, corn steep liquor, casein. Hydrolyzate, soybean meal and soybean meal hydrolyzate, various fermented microorganisms and digested products thereof, etc. can be used.

As the inorganic salt, for example, potassium phosphate dibasic, potassium phosphate dibasic, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate, and/or calcium carbonate can be used.

The culture is preferably performed under aerobic conditions such as shaking culture or deep aeration stirring culture. The culture temperature is preferably 15 to 40°C, and the culture time is usually preferably 16 hours to 7 days. The pH during culture is preferably maintained at 3.0 to 9.0. Adjustment of pH can be performed using an inorganic or organic acid, an alkali solution, urea, calcium carbonate, ammonia, etc.

In addition, an antibiotic such as ampicillin or tetracycline may be added to the medium during culture as needed.

As a medium for culturing transformants obtained by using insect cells as hosts, commonly used TNM-FH medium (manufactured by Becton Dickinson), Sf-900 II SFM medium (manufactured by Invitrogen), ExCell400, ExCell405 (all of which are JRH Biosciences Co.), Grace's insect medium (Nature, 195, 788 (1962)), or Schneider's Medium (Thermo Fisher) can be used.

The culture of the transformant obtained using insect cells as a host is usually preferably performed at a pH of 6 to 7 and a temperature of 25 to 30°C, preferably for 1 to 5 days. In addition, you may add an antibiotic substance, such as gentamicin, to a medium as needed during culture|cultivation.

As the medium for animal cell culture, RPMI1640 medium (manufactured by Invitrogen), GIT medium (manufactured by Nippon Seiyaku), EX-CELL301 medium (manufactured by JRH), EX-CELL325 PF CHO serum-free medium (Sigma Aldlich), IMDM Medium (manufactured by Invitrogen), Hybridoma-SFM medium (manufactured by Invitrogen), Eagle's minimal essential medium (MEM) [Science, 122, 501 (1952)], Dulbecco's modified Eagle's medium [Virology, 8, 396] (1959)], 199 medium [Proceeding of the Society for the Biological Medicine, 73, 1 (1950)], or a medium in which various additives such as FBS are added to these medium are used.

Cultivation of animal cells is usually preferably performed in the presence of 5% CO ₂ , preferably at a pH of 6 to 8 and a temperature of 30 to 40° C., and preferably for 1 to 7 days. In addition, you may add an antibiotic substance, such as kanamycin or penicillin, to a culture medium as needed during culture.

By culturing the obtained transformant in a medium, the expression and accumulation of the DcR3 variant or the DcR3 variant in the culture supernatant. In addition, the transformant can increase the expression level of a DcR3 variant or a DcR3 variant using a DHFR amplification system (Japanese Patent Application Laid-Open No. 2-257891) or the like.

By producing and accumulating the DcR3 variant according to the present invention in the culture as described above, and collecting from the culture, the DcR3 variant according to the present invention can be produced.

In order to isolate and purify the peptide produced by the transformant, a conventional protein isolation and purification method can be used.

For example, when the DcR3 variant of the present invention is secreted extracellularly, the DcR3 variant can be recovered from the culture supernatant. That is, a culture supernatant is obtained by treating the culture by a method such as centrifugation, and from the culture supernatant, a normal protein isolation and purification method, that is, a solvent extraction method, a salting-out method with ammonium sulfate, etc., a desalting method, an organic solvent precipitation method using a precipitation method, anion exchange chromatography method using a resin such as diethylaminoethyl (DEAE)-Sepharose, or DIAION HPA-75 (Mitsubishi Chemical Corporation), S-Sepharose FF (GE Healthcare Corporation), etc. Cation exchange chromatography using a resin, hydrophobic chromatography using a resin such as butyl sepharose or phenyl sepharose, gel filtration using molecular sieves, affinity chromatography, chromatographic focusing, or isoelectric electrophoresis A refined product can be obtained by using methods, such as an electrophoresis method, such as these alone or in combination.

For example, when the DcR3 variant of the present invention has an immunoglobulin Fc capable of binding to protein G or protein A, protein G or protein A is bound to a carrier as an affinity ligand, protein G chromatography or protein A A chromatography method can be used as an affinity chromatography method (Monoclonal Antibodies-Principles and practice, Third edition, Academic Press (1996), Antibodies-A Laboratory Manual, Cold Spring Harbor Laboratory (1988)). In addition, methods used in protein purification, such as gel filtration, ion exchange chromatography, and ultrafiltration, may be combined.

In addition, by using a chemical modification method, which is a conventional method, from the DcR3 modified product obtained by the above method, it is also possible to obtain a DcR3 modified product further chemically modified with a peptide, sugar chain, or PEG.

6. Method of evaluation of biological activity, physical properties and kinetics of DcR3 variants

As the DcR3 variant of the present invention, preferably LIGHT, TL1A and DcR3 variant having neutralizing activity with respect to at least one or more of FasL, LIGHT, DcR3 variant having neutralizing activity for all of TL1A and FasL, FasL DcR3 variants that do not have neutralizing activity, and have neutralizing activity against either LIGHT and TL1A, and DcR3 variants that do not have neutralizing activity against FasL and have neutralizing activity against LIGHT and TL1A, and the like. .

The DcR3 variant of the present invention can measure biological activity such as neutralizing activity, physical properties and in vivo dynamics by using the following method.

(1) Preparation of Ligand

LIGHT, TL1A, and FasL used in the present invention include, but are not limited to, LIGHT, TL1A and FasL derived from eukaryotes. Examples of the eukaryotic-derived LIGHT, TL1A and FasL include yeast, insects, and mammals. Preferably, LIGHT, TL1A, and FasL derived from a primate including a human or a rodent including a mouse are mentioned.

For cells expressing LIGHT, TL1A or FasL or the ligand, an expression vector containing cDNA encoding the full length of LIGHT, TL1A or FasL or a partial length thereof, Escherichia coli, yeast, insect cells, or animal cells, etc. is suitable It can be obtained by introducing into a host cell. It can also be obtained by purifying the ligand from various human cultured cells or human tissues expressing large amounts of LIGHT, TL1A or FasL. In addition, the cultured cells, the tissue, or the like can also be used as it is as a ligand. In addition, a synthetic peptide having a partial sequence of LIGHT, TL1A or FasL can also be prepared by a chemical synthesis method such as the Fmoc method or the tBoc method.

In addition, LIGHT, TL1A and FasL use the method described in Molecular Cloning, A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press (1989), etc., for example, by the following method, LIGHT, TL1A or FasL It can be prepared by introducing and expressing the encoding DNA into a host cell.

Soluble LIGHT, TL1A, and FasL are produced by expressing on the cell membrane as membrane LIGHT, TL1A, and FasL, respectively, and then shedding the extracellular region with a protease. The cleavage site has been identified for each ligand, but any sequence in the vicinity of the cleavage site may be included when a recombinant of a soluble ligand is prepared. For example, a soluble LIGHT recombinant can be prepared from the region 66 to 240, the 74 to 240 region, the 83 to 240 region from the N-terminus in the amino acid sequence of the membrane LIGHT. A soluble TL1A recombinant can be prepared from, for example, the region 72 to 251 from the N-terminus in the amino acid sequence of membrane-type TL1A. Soluble FasL can be prepared from the region 130 to 281, the region 134 to 281, or the like from the N-terminus in the amino acid sequence of membrane FasL.

A recombinant of a soluble ligand can be prepared by adding a His tag, a FLAG tag, or the like to the N-terminus of the region, and can be obtained by affinity purification. Examples of amino acid sequences of LIGHT, TL1A and FasL soluble recombinants include amino acid sequences set forth in SEQ ID NOs: 114, 116, 118, 120, 122, 124, 126, 128, 130 and 306. As the nucleotide sequence of the DNA encoding the amino acid sequence of the LIGHT, TL1A and FasL soluble recombinants, for example, the nucleotide sequences shown in SEQ ID NOs: 113, 115, 117, 119, 121, 123, 125, 127, 129 and 305 can be heard

Functional LIGHT, TL1A or FasL can form homo trimers. The molecular weight can be calculated|required by SEC-MALS analysis of the soluble LIGHT, TL1A, or FasL prepared by any of the methods mentioned above. SEC-MALS refers to the scattering intensity detected by Multi Angle Light Scattering (MALS) for each peak detected at a wavelength of 215 nm separated by SEC (Size Exclusion Chromatography)-HPLC. It is an analysis method that calculates the molecular weight using the maximum value. As an HPLC system, For example, Shimadzu Corporation's Prominence, as an SEC column, For example, TSKgel of Tosoh Corporation, as a detector of MALS, Wyatt Technology's miniDAWN TREOS etc. are mentioned, for example. Moreover, as a method of isolating a homo trimer from the crude substance which forms other than a trimer, SEC purification etc. are mentioned, for example. As a method of SEC purification, for example, using AKTApurifier manufactured by GE Healthcare as an HPLC system and Superdex 200 Increase 10/300 GL manufactured by GE Healthcare as an SEC column, the crude product is fractionated according to molecular weight size, and homo trimer a method of recovering only the molecular weight fraction of

(2) Evaluation of binding activity of DcR3 variants comprising a human Fc region

As a method of measuring the binding activity of the DcR3 variant to soluble LIGHT, soluble TL1A or soluble FasL, for example, binding analysis by enzyme immunoassay (ELISA) and kinetics analysis by Biacore, etc. can be heard As the ligand, a transgenic cell or recombinant protein obtained by introducing an expression vector containing a DNA sequence encoding the extracellular domain of each ligand of LIGHT, TL1A or FasL to E. coli, yeast, insect cells or animal cells, and human tissue or Serum, plasma, or culture supernatant containing a ligand obtained from human cells is used.

In the case of ELISA, for example, an anti-human IgG antibody is immobilized on a 96-well plate or the like, reacted with a DcR3 variant, and then each ligand is dispensed and bound. After washing, an unlabeled anti-ligand antibody or a receptor-Fc fusion of each ligand is reacted, followed by reaction with an anti-Fc antibody labeled with biotin, an enzyme, or a chemiluminescent substance, or a labeled anti-ligand antibody or Binding of the DcR3 variant to the ligand can be measured by reacting each ligand with a receptor-Fc fusion or the like, and performing detection according to a labeling substance. In the case of Biacore, for example, using Biacore T100 or Biacore T200, the kinetics in the binding of each ligand to the DcR3 variant is measured, and the result is analyzed with the analysis software provided with the device. Specifically, after immobilizing an anti-human IgG antibody to the sensor chip CM5 by amine coupling, a DcR3 variant is flowed to bind to the sensor chip in an appropriate amount, and a plurality of ligands of known concentrations are further flowed to bind, measure the dissociation. The obtained data is subjected to kinetic analysis by a 1:1 binding model using the software attached to the device, and various parameters are acquired. Alternatively, after each ligand is immobilized on a sensor chip by, for example, an amine coupling method, a plurality of DcR3 variants of known concentrations are flowed to measure binding and dissociation. The obtained data is kinetically analyzed by the bivalent binding model using the software attached to the device, and various parameters are acquired. Alternatively, the DcR3 variant is flowed to the sensor chip protein A in which the MabSelectSure ligand, which is an IgG-binding domain variant protein of protein A, is immobilized on the sensor chip, and an appropriate amount is bound to the sensor chip, and a plurality of ligands of known concentrations are further flowed. Thus, binding and dissociation are measured. The obtained data is subjected to kinetic analysis by a 1:1 binding model using the software attached to the device, and various parameters are acquired.

The binding activity of the DcR3 variant to membrane-type LIGHT, membrane-type TL1A or membrane-type FasL is, for example, a gene obtained by introducing an expression vector containing a DNA sequence encoding the full length of each ligand of LIGHT, TL1A and FasL to animal cells, etc. Mitogen stimulation, such as PHA-L, PMA, or Ionomycin, or anti-CD3 antibody and anti-CD28 antibody stimulation, cytokine, IgG, or immunity to transduced cells or human cells such as PBMCs or HUVECs It can be measured by flow cytometry using cells in which expression of a membrane ligand is induced by applying an appropriate stimulus such as complex stimulation. Specifically, the DcR3 variant is reacted with cells expressing the membrane ligand, reacted with a fluorescently-labeled anti-Fc antibody, or the like, or the DcR3 variant labeled with biotin or a fluorescent dye, etc. is reacted. After washing, flow It can be measured by using a cytometer to detect the fluorescence intensity according to the labeling substance.

As measured by the above-described method, the DcR3 variant of the present invention, for example, LIGHT, TL1A, and DcR3 variant having binding activity to at least one or more of FasL, LIGHT, TL1A and FasL having binding activity to all of DcR3 variants, without binding activity to FasL, and having no binding activity to either LIGHT and TL1A, or DcR3 variants having binding activity to FasL, and having binding activity to LIGHT and TL1A DcR3 variants can be mentioned.

(3) Evaluation of neutralizing activity of DcR3 variants

As a method for measuring the neutralizing activity of the DcR3 variant, for example, in a solution to which the DcR3 variant is added, a method for measuring the binding of LIGHT, TL1A or FasL to the corresponding receptor, or in a medium to which the DcR3 variant is added In this case, there is a method in which a corresponding ligand is added to LIGHT, TL1A or FasL receptor-expressing cells to measure cellular functions such as cytokine production and cell proliferation.

The inhibitory activity of human LIGHT on binding to a corresponding receptor can be measured by the following method, for example, using the method described in US Pat. No. 8,974,787B2. In the reaction solution to which the DcR3 variant was added, HVEM or LTβR labeled with biotin or a fluorescent dye was reacted with cells expressing membrane-type LIGHT, and after washing, the fluorescence intensity according to the labeling substance was measured using a flow cytometer. It is measured by detection. Alternatively, in the reaction solution to which the DcR3 variant is added, LIGHT labeled with biotin or a fluorescent dye is reacted with cells expressing HVEM or LTβR, and after washing, the fluorescence intensity according to the labeling substance using a flow cytometer It is measured by detecting The inhibitory activity of the DcR3 variant with respect to the binding of LIGHT to the corresponding receptor is confirmed by the decrease in the fluorescence intensity, in comparison with the case where the DcR3 variant is not added.

Inhibitory activity for binding of TL1A or FasL to the corresponding receptor can also be measured using the method similar to the above.

As the DcR3 variant of the present invention, LIGHT, TL1A and DcR3 variant having inhibitory activity with respect to at least one or more of the binding of the corresponding receptor to FasL, LIGHT, TL1A and FasL for all binding of the corresponding receptor DcR3 variant having inhibitory activity, DcR3 variant having no inhibitory activity with respect to the binding of FasL and the corresponding receptor, and also having inhibitory activity with respect to any one of the binding of LIGHT and TL1A to the corresponding receptor, or FasL and A DcR3 variant having no inhibitory activity with respect to the binding of the corresponding receptor and also having inhibitory activity with respect to the binding of LIGHT and TL1A to the corresponding receptor is preferred. In the present invention, the expression "having no inhibitory activity with respect to the binding of a certain ligand to a corresponding receptor" is used in the meaning including that the inhibitory activity is significantly lower than that of wild-type DcR3.

The inhibitory activity with respect to the cell function caused by the addition of LIGHT can be measured as follows, using the method etc. described in US Patent US8,974,787B2, for example. In the medium to which the DcR3 variant is added, LIGHT-dependent IL-8, CCL5 or CCL20, etc. using cells such as HT-29 (ATCC number: HTB-38), which have been confirmed to express LIGHT receptor HVEM or LTβR, etc. chemokine production is measured by an ELISA method using a culture supernatant, alphaLISA (Perkin Elmer), or CBA analysis (BD Biosciences) or the like. Alternatively, using cells such as long myofibroblasts (Lonza) stimulated with IFN-γ, soluble or membranous LIGHT-dependent chemokine production such as CXCL-10 is measured using the same method. Alternatively, instead of soluble or membrane-type LIGHT, anti-CD3 antibody, anti-CD28 antibody, or LIGHT expression induced from PBMCs or T cells stimulated with PMA and ionomycin or the like can be used for measurement in the same manner. The inhibitory activity of the DcR3 variant on cellular function caused by the addition of soluble or membrane LIGHT is confirmed by the decrease in the amount of production of the chemokine in comparison with the case where the DcR3 variant is not added. In vivo, for example, in an acute GVHD (Graft versus host disease) model in which human PBMCs were trans-transplanted into immunocompromised mice using the method described in US Pat. No. 8,974,787 B2, etc. , it is possible to evaluate the improvement of survival, body weight, condition, pathology, engraftment of human cells, etc. by administration of the DcR3 variant.

The inhibitory activity on cell function caused by the addition of TL1A can be measured as follows, for example, using the method described in Mucosal Immunology, 2015, 8: p.545-558. In the medium to which the DcR3 variant is added, using blood, PBMC, pan T cells, CD4-positive T cells, or memory CD4-positive T cells derived from primates such as humans or rodents such as mice, IL-12, IL -18 and TL1A, or each cytokine cocktail of IL-12, IL-18, IL-15 and TL1A stimulated the corresponding cells, TL1A-dependent IFN-γ, IL-6, GM-CSF, TNF-α, The production of cytokines such as IL-5, IL-13, IL-17 or IL-22 is measured by ELISA method using culture supernatant, alphaLISA or CBA analysis, or the like. Alternatively, for example, using the method described in Immunity, 2002, 16: p.479-492, in a medium to which a DcR3 variant is added, pan T cells derived from primates such as humans or rodents such as mice, CD4 Positive T cells or memory CD4 positive T cells are stimulated with an anti-CD3 antibody or an anti-CD28 antibody, and TL1A-dependent IFN-γ and IL-2 production is measured by the same method as described above. Alternatively, measurement can also be carried out in a similar manner using a forced expression strain of membrane TL1A instead of soluble TL1A. Alternatively, the measurement can also be carried out in the same manner using PBMCs stimulated with IgG or immune complexes or TL1A expression induced from monocytes. The inhibitory activity of the DcR3 variant on cellular function caused by the addition of TL1A is confirmed by the decrease in the cytokine or inhibition of cell proliferation in comparison with the case where the DcR3 variant is not added. Further, in vivo, for example, the TNBS (2,4,6-trinitrobenzenesulfonic acid)-induced colitis model described in Mucosal Immunology, 2011, 4: p.172-185, or Mucosal Immunology, 2014, 7: p.1492 In the DSS (dextran sodium sulfate)-induced colitis model described in -1503, improvement in survival, body weight, condition and pathology by administration of a DcR3 variant can be evaluated. In addition, the pathological improvement effect in TL1A-dependent inflammation, allergic disease, or autoimmune disease models in rodents such as mice can be evaluated.

The inhibitory activity with respect to the cell function caused by the addition of FasL can be measured as follows, for example, using the method described in J. Rheumatol., 2013, 40: p.1316-1326. In the medium to which the DcR3 variant was added, Jurkat cells (DSMZ number: ACC282) were used to induce soluble FasL or soluble FasL-dependent apoptosis cross-linked with an antibody, by staining with Annexin V/Propium Iodide or introducing BrdU into live cells. It is measured by the amount of ATP, etc. Alternatively, measurement can also be carried out in a similar manner using a membrane-type FasL forced expression strain instead of soluble FasL. Alternatively, it can be measured in the same way using FasL derived from stimulated PBMCs or T cells. The inhibitory activity of the DcR3 variant on cellular function caused by the addition of FasL is confirmed by the reduction in apoptosis compared to the case where the DcR3 variant is not added.

As the DcR3 variant of the present invention measured by the above-described method, for example, LIGHT, TL1A and DcR3 variant having inhibitory activity against at least one or more of the cellular functions caused by the addition of FasL, LIGHT, TL1A and FasL DcR3 variant having inhibitory activity against all of the cellular functions caused by the addition of FasL does not have inhibitory activity on the cellular functions caused by the addition of FasL, and among the cellular functions caused by the addition of LIGHT and TL1A DcR3 variants having inhibitory activity against any one or more, or DcR3 having no inhibitory activity on cellular functions caused by the addition of FasL, and also having inhibitory activity on cellular functions caused by the addition of LIGHT and TL1A A modified body can be mentioned. In the present invention, the expression "having no inhibitory activity with respect to the cellular function caused by the addition of a certain ligand" is used in the meaning including that the inhibitory activity is significantly lower than that of wild-type DcR3.

(4) OPG ligand reactivity evaluation of DcR3 variants

One of the features of the DcR3 variant of the present invention is that any or all of CRD1, CRD4, and CRD3 parts that are not involved in ligand binding are OPG-derived sequences.

The fact that the DcR3 variant does not have binding activity to OPG ligands RANKL and TRAIL can be evaluated, for example, by the same method as the method for measuring the binding activity to LIGHT, TL1A and FasL described above.

One of the features of the DcR3 variant of the present invention is that it does not have neutralizing activity also with respect to RANKL and TRAIL.

The neutralizing activity of the DcR3 variant on RANKL is, for example, described in J. Immunol., 2012, 189: p.245-252, TRAP (tartrate-resistant acid phosphata It can be evaluated using tartrate-resistant acid phosphatase activity measurement or the like.

In addition, the neutralizing activity of the DcR3 variant with respect to TRAIL, for example, in a human cancer cell line expressing DR4 or DR5, apoptosis induction by soluble TRAIL or cross-linked soluble TRAIL, neutralizing activity against FasL described above It can be evaluated by the method similar to the method of measuring .

(5) Dynamic evaluation of DcR3 variants

As the DcR3 variant of the present invention, a DcR3 variant having reduced or lost binding to heparan sulfate contained in heparan sulfate proteoglycan (HSPG) on the cell membrane is preferable, and one that does not have a heparan sulfate binding domain (HBD) is preferred. more preferably.

The presence or absence of binding to heparan sulfate on the cell membrane via HBD can be measured as follows, for example, using the method described in J. Immunol., 2006, 176: p.173-180. Any cells, for example, CHO cells, human cell lines, vascular endothelial cells, hepatocytes, or blood cells, etc. are reacted with wild-type DcR3 or a DcR3 variant, followed by reaction with a fluorescently-labeled detection antibody, or the like, biotin, a fluorescent dye. It can be measured by reacting wild-type DcR3 or a DcR3 mutant labeled with, etc., washing, and detecting the fluorescence intensity according to the labeling material using a flow cytometer (FCM). The decrease or loss of DcR3 binding to the cell membrane due to the deletion of HBD can be investigated by the decrease in fluorescence intensity by FCM.

As a method of evaluating whether the binding to the cell membrane is binding of DcR3 through HBD, for example, using the method described in J. Immunol., 2006, 176: p.173-180, wild-type DcR3 or DcR3 Examples of a method of reacting wild-type DcR3 or DcR3 variant by adding GAG such as heparin or heparan sulfate as an inhibitor during the reaction of the variant, or by pretreating the cells with an enzyme such as heparinase or trypsin, etc. can be heard as

One of the features of the modified DcR3 of the present invention is that its in vivo kinetics is improved compared to that of wild-type DcR3 because its loss through heparan sulfate is reduced in vivo. In the present invention, "improved in vivo kinetics" means that the blood half-life is longer than that of wild-type DcR3, or the area under the concentration-time curve (AUC) value up to infinity is high. do.

In addition, as a comparison object, a molecule containing the CRD of wild-type DcR3 may be used instead of wild-type DcR3. Specifically, DcR3 FL-Fc (amino acid sequence: SEQ ID NO: 100, DNA nucleotide sequence: SEQ ID NO: 99), DcR3 FL-FLAG (amino acid sequence: SEQ ID NO: 104, DNA nucleotide sequence: SEQ ID NO: 103), wild-type DcR3 S195-Fc (amino acid sequence: SEQ ID NO: 102, DNA nucleotide sequence: SEQ ID NO: 101) and full-length DcR3 having 1 amino acid mutation in HBD (amino acid sequence: SEQ ID NO: 112, DNA nucleotide sequence) : R128Q-Fc in which Fc is fused to the C terminus of SEQ ID NO: 111) [US Pat. No. 6,835,814 B1, US6,965,01 B1], and the like. When a molecule comprising the CRD of these wild-type DcR3 is used as a comparison object, it is also referred to as a wild-type DcR3 control.

In rodents such as mice, and non-human primates such as cynomolgus monkeys, the change in blood concentration of the DcR3 variant is determined by administering an arbitrary dose intravenously or subcutaneously, collecting blood at any time, and using a detection system for DcR3 or human Fc. can be used to measure Kinetic parameters such as blood half-life and AUC can be calculated from the blood concentration transition using methods such as moment analysis using, for example, the method described in Pharmacokinetics, 1999, 14: p.286-293. .

In addition, since it is known that in vivo dynamics is improved when the number of sialic acids added to the terminal of the N-type sugar chain is large (J. Pharm. Sci., 2015, 104: p.1866-1884), the present invention having an N-type sugar chain In the DcR3 variant, it is also preferable that the number of additions of sialic acid is large. The number of additions of sialic acid per protein molecule is calculated by separating the labeled sialic acid by reverse-phase HPLC using, for example, a reagent kit for sialic acid fluorescent labeling (Takara), etc., and comparing it with a standard curve of sialic acid. can do.

(6) Evaluation of physical properties of the DcR3 variant

When the DcR3 variant of the present invention is expressed, isolated or purified using mammalian cells, one feature is that the content of aggregates is lower than that of wild-type DcR3.

Whether or not aggregates are generated during protein expression and secretion, for example, by recovering a host cell and/or culture supernatant into which a recombinant vector carrying a DcR3 variant is introduced, anti-DcR3 antibody, or Fc fusion or His; In the case of tag adducts such as FLAG, it can be judged by examining the approximate molecular weight under non-reducing conditions by immunoblotting with an antibody against them. When proteins are aggregated during protein expression and/or secretion, one or more bands having a size larger than the expected molecular weight are detected.

The method of calculating the molecular weight of a protein from an amino acid sequence is mentioned. Moreover, when calculating|requiring a more accurate molecular weight, the analysis method by SEC-MALS is mentioned. For each peak detected at a wavelength of 215 nm separated by SEC (size exclusion chromatography)-HPLC, the molecular weight can be calculated using the maximum value of the scattering intensity detected by a Multi Angle Light Scattering (MALS). . As an HPLC system, For example, Shimadzu Corporation's Prominence, as an SEC column, For example, TSKgel of Tosoh Corporation, as a detector of MALS, Wyatt Technology's miniDAWN TREOS etc. are mentioned, for example.

Whether the isolated or purified protein is aggregated can be detected by SDS-PAGE of the protein under non-reducing conditions, CBB staining or the like, or by immunoblotting in the same manner as described above. In addition, as for the content of aggregates, with respect to each peak detected at a wavelength of 215 nm separated by gel filtration chromatography (SEC) using HPLC, the ratio of each peak can be calculated from the area. As an SEC column, TSKgel G3000SW by Tosoh Corporation, ACQUITY UPLC Protein BEH SEC by Waters, etc. are mentioned, for example. In the case of analysis by the above-described method, the preferred content of aggregates is 0 to 60%, the more preferred content of aggregates is 0 to 40%, the more preferred content of aggregates is 0 to 30%, and the more preferred content of aggregates is 0 to 30%. The content is 0 to 20%, and the most preferred agglomerate content is 0 to 10%.

In addition, the DcR3 variant of the present invention is characterized in that the hydrophobicity is lower than that of wild-type DcR3.

The hydrophobicity of a protein may be measured using a hydrophobic interaction chromatography (HIC) column that interacts with a hydrophobic region existing on the surface of the protein. As an HIC column, TSKgel Butyl-NPR by Tosoh Corporation etc. are mentioned, for example.

In addition, the DcR3 variant of the present invention is characterized in that the thermal stability is improved than that of wild-type DcR3. As a method for measuring the thermal stability of a protein, a calorimetry method such as DSC (Differential scanning calorimetry), autofluorescence spectrum or circular dichorism (CD) at the time of thermal denaturation or chemical denaturation Spectroscopy for obtaining a spectrum, DSF (differential scanning fluorescence system) detecting exposure of a hydrophobic region present inside a protein with a temperature rise with a fluorescent dye (Sypro Orange, etc.) [J Am Chem Soc, 2009, 131: p. 3794-3795].

DSF for evaluating the thermostability of proteins is described, for example, in J. Pharm. Sci., 2013, 102: The amount of fluorescence at each temperature can be measured using the method etc. described in p.2471-2483. In addition, using software such as BioRad's CFX manager, a melting curve can be drawn and a Tm (thermal unfolding transition midpoints) value can be calculated. Likewise, DSC for evaluating the thermostability of proteins is described, for example, in J. Pharm. Sci., 2012, 101: The heat capacity and Tm value in each temperature can be computed using the method etc. described in p.955-964.

7. DcR3 variant composition

Examples of the modified DcR3 composition of the present invention include a composition containing a modified DcR3 molecule. In the DcR3 variant composition of the present invention, the primary amino acid sequences constituting the DcR3 variant molecule are equivalent, and post-translational modifications such as oxidation/reduction reaction, sugar chain addition reaction, sulfate addition reaction, etc. in the amino acid sequence and a composition containing a plurality of DcR3 variant molecules that may occur. The DcR3 variant of the present invention may include, for example, a DcR3 variant having one or more N-glycosidic conjugated sugar chains and a DcR3 variant having no N-glycosidic conjugated sugar chain. The ratio of the DcR3 variants having one or more N-glycosidic conjugated sugar chains is, for example, preferably 70 to 100%, more preferably 90 to 99%, with respect to the total number of DcR3 variants of the present invention. and particularly preferably 95 to 98%.

8. Pharmaceutical composition comprising a DcR3 variant

One embodiment of the present invention is a composition comprising an effective amount of the DcR3 variant of the present invention. The composition comprising the modified DcR3 variant of the present invention can be used as an active ingredient for a prophylactic or therapeutic agent for autoimmune diseases, inflammatory diseases, or allergic diseases, including mucosal diseases. That is, by administering a pharmaceutical composition comprising the DcR3 variant of the present invention to a patient in need of prevention or treatment of an autoimmune disease, an inflammatory disease, or an allergic disease, the autoimmune disease, an inflammatory disease or an allergic disease can be prevented or treated can

Conditions or diseases in which the composition of the present invention is used include, but are not limited to, for example, inflammatory bowel disease (IBD), systemic erythematosus, psoriasis, chronic graft-versus-host disease, acute graft-versus-host disease, Crohn's disease, Ulcerative colitis, inflammatory bowel disease, multiple sclerosis, celiac disease, idiopathic thrombotic thrombocytopenic purpura, myasthenia gravis, Sigren's syndrome, scleroderma, asthma, uveitis, epidermal hyperplasia, alopecia areata, Behcet's disease, Takayasu's arteritis, cartilage Inflammation, osteolysis, arthritis, juvenile arthritis, juvenile joint rheumatism, microarticular juvenile joint rheumatism, polyarticular juvenile joint rheumatism, systemic onset juvenile joint rheumatism, juvenile ankylosing spondylitis, juvenile Enteropathic arthritis, juvenile reactive arthritis, juvenile Reiter's syndrome, SEA syndrome (seronegative, tendonitis, arthrosis syndrome), juvenile dermatomyositis, juvenile psoriatic arthritis, juvenile scleroderma, juvenile systemic erythema Todes, juvenile vasculitis, microarticular rheumatism, polyarticular rheumatoid arthritis, generalized aplastic joint rheumatism, ankylosing spondylitis, enteropathic arthritis, reactive arthritis, Reiter's syndrome, dermatomyositis, psoriatic arthritis, vasculitis, myositis , polymyositis, dermatomyositis, osteoarthritis, polyarteritis nodosa, Wegener's granuloma, arteritis, polymyalgia rheumatica, sarcoidosis, sclerosis, primary biliary cirrhosis, sclerosing cholangitis, dermatitis, atopic dermatitis, atherosclerosis, Inflammatory diseases such as Still's disease, chronic obstructive pulmonary disease, Guillain-Barré syndrome, type 1 diabetes, Graves disease, Addison's disease, Raynaud's phenomenon, autoimmune hepatitis, or Wiscott-Aldrich syndrome, autoimmune diseases, or allergic diseases and the like.

The pharmaceutical composition containing the modified DcR3 of the present invention may contain a mixture with the modified DcR3 or any other therapeutic active ingredient as an active ingredient. In addition, these pharmaceutical preparations are prepared by mixing the active ingredient with one or more pharmacologically acceptable carriers, and any method well known in the art of pharmaceuticals.

The content of the modified DcR3 of the present invention in the pharmaceutical composition varies depending on the dosage form and the pharmacologically acceptable dosage of the modified DcR3 of the present invention, but is, for example, about 0.01 to 100% by weight. In addition, the content of the pharmacologically acceptable carrier in the pharmaceutical preparation varies depending on the dosage form, the pharmacologically acceptable dose of the DcR3 modified body of the present invention, and the like, but is, for example, 0 to 99.9% by weight.

As the route of administration, it is preferable to use the one most effective for treatment, for example, oral administration, or parenteral administration such as intravenous, subcutaneous, oral, airway, rectal, intramuscular, or intraperitoneal. can

The dosage form includes tablets, powders, granules, syrups or injections.

Suitable preparations for oral administration include, for example, liquid preparations such as syrups, saccharides such as water, sucrose, sorbit, or fructose, glycols such as polyethylene glycol or propylene glycol, sesame oil, olive oil, or soybean oil. It can be manufactured using oils, preservatives such as p-hydroxybenzoic acid esters, strawberry flavor, or flavors such as peppermint. In addition, tablets, powders and granules, etc., are excipients such as lactose, glucose, sucrose, or mannite, disintegrants such as starch or sodium alginate, lubricants such as magnesium stearate or talc, polyvinyl alcohol, hydroxy It can be prepared by using a binder such as propyl cellulose or gelatin, a surfactant such as fatty acid ester, and a plasticizer such as glycerin.

Formulations suitable for parenteral administration include sterile aqueous preparations containing the active compound, preferably isotonic with the blood of the recipient. For example, in the case of an injection, a solution for injection is prepared using a carrier including a salt solution, a glucose solution, or a mixture of saline and glucose solution.

In addition, also in these parenteral preparations, one or more auxiliary ingredients selected from diluents, preservatives, flavors, excipients, disintegrants, lubricants, binders, surfactants and plasticizers exemplified in oral preparations may be added. .

The drug containing the DcR3 variant of the present invention can be safely administered to mammals (eg, humans, mice, rats, rabbits, dogs, cats, cattle, horses, pigs, monkeys, etc.).

The dosage and frequency of administration of the DcR3 variant of the present invention vary depending on the dosage form, the age, weight, disease, and the nature or severity of the symptom to be treated, but in the case of oral administration, 0.01 mg to 1 g per adult, Preferably, 0.05 to 50 mg is administered once to several times a day. In the case of parenteral administration such as intravenous administration, 0.001 to 100 mg, preferably 0.01 to 10 mg, per adult is administered once to several times a day. However, these dosages and the frequency of administration vary depending on the various conditions described above.

Example

Examples of the present invention are shown below. However, the present invention is not limited by these examples.

[Example 1] Evaluation of aggregation of wild-type DcR3 in mammalian cells

Fusion protein (DcR3 FL-Fc) of wild-type DcR3 (also called full-length DcR3) (SEQ ID NO: 4) with linker sequence IEGRMD (SEQ ID NO: 106) and human IgG1 Fc region (g1S) (SEQ ID NO: 72) ( FIG. 3 ) A, SEQ ID NO: 100), a fusion protein (S195-Fc) of a human DcR3 (SEQ ID NO: 108) lacking the HBD region, a linker sequence IEGRMD (SEQ ID NO: 106), and a human IgG1 Fc region (SEQ ID NO: 72) ( 3B, SEQ ID NO: 102), and a protein (DcR3 FL-FLAG) (SEQ ID NO: 104) in which a FLAG tag (SEQ ID NO: 110) was added to full-length DcR3 (SEQ ID NO: 4) was constructed as follows.

For DcR3 FL-Fc, the DNA fragment of the signal peptide sequence, the DNA fragment of human DcR3 (SEQ ID NO: 3), the DNA fragment of the linker sequence IEGRMD (SEQ ID NO: 105), the DNA fragment of Fc (g1S) (SEQ ID NO: 71) Inserted using the In-Fusion HD Cloning Kit (Clontech) into pCIpuro vector (partially modified from Promega's pCI) artificially synthesized (GENEWIZ or Sigma) and digested with restriction enzymes NheI and SalI (New England Biolabs) Then, Escherichia coli DH5α competent cells (TOYOBO) were transformed to obtain a transformant into which the DcR3 FL-Fc DNA fragment (SEQ ID NO: 99) was inserted. Similarly for S195-Fc, a DNA fragment of a signal peptide sequence, a DNA fragment of human DcR3 excluding HBD (SEQ ID NO: 107), a DNA fragment of a linker sequence IEGRMD (SEQ ID NO: 105), a DNA fragment of Fc (g1S) (SEQ ID NO: 71) was artificially synthesized to obtain a transformant into which the S195-Fc DNA fragment (SEQ ID NO: 101) was inserted. For DcR3 FL-FLAG, a DNA fragment of the signal peptide sequence, a DNA fragment of human DcR3 (SEQ ID NO: 3), and a DNA fragment of a FLAG tag (SEQ ID NO: 109) were artificially synthesized, and the DcR3 FL-FLAG DNA fragment (SEQ ID NO: 103) ) to obtain an inserted transformant.

Each plasmid obtained from each transformant was introduced into any host cell of Freestyle CHO-S cells, Freestyle 293F cells, and Expi293 cells (all manufactured by Thermo Scientific) to transiently express proteins. For plasmid introduction, any of Freestyle MAX Reagent, 293Fectin Transfection Reagent, and ExpiFectamine 293 (all manufactured by Thermo Scientific) was used.

After culturing the transfected cells for several days, the culture supernatant was recovered, and DcR3 FL-Fc and S195-Fc were used using MabSelect SuRe (GE Healthcare), and also Anti-FLAG M2 affinity gel (Sigma) was used. and DcR3 FL-FLAG were each affinity-purified. The culture supernatant of DcR3 FL-Fc and S195-Fc was passed through a column filled with resin, washed with PBS (Nacalai Tesque), and eluted with an elution buffer (20 mM citric acid, 50 mM NaCl, pH3.4), It was quickly neutralized with a neutralization buffer (1 M Na phosphate, pH 7.0). The culture supernatant of DcR3 FL-FLAG was similarly filled with resin, washed with PBS, eluted with an elution buffer (100 mM glycine-HCl, pH3.5), and rapidly neutralized with a buffer for neutralization (1M Tris-HCl, pH8.0) was neutralized with The absorbance (A ₂₈₀ ) at 280 nm of each elution fraction was measured, and continuous fractions with high measured values were collected. The buffer solution of the recovered fraction was replaced with PBS using a NAP column (GE Healthcare) and passed through a 0.22 µm filter as purified protein. Concentrations were calculated with the extinction coefficients of DcR3 FL-Fc, S195-Fc and DcR3 FL-FLAG at 280 nm being 1.03, 1.17, and 0.77, respectively, and after SDS-PAGE under non-reducing and reducing conditions of 100 mM DTT, The gel was subjected to Coomassie staining (Nacalai Tesque) to confirm the molecular weight. The estimated molecular weights predicted from the amino acid sequences of DcR3 FL-Fc, S195-Fc and DcR3 FL-FLAG are about 56.4 kDa, 44.7 kDa, and 31.0 kDa in the monomer, respectively. Under non-reducing conditions, the Fc fusions, DcR3 FL-Fc and S195-Fc, exist as dimers, so that their estimated molecular weights are about 112.8 kDa and 89.4 kDa, respectively.

As a result of SDS-PAGE under non-reducing conditions, most of DcR3 FL-Fc, S195-Fc, and DcR3 FL-FLAG transiently expressed in mammalian cells remained in the sample wells, and also those that were migrated were smeared or It became a ladder shape, and it became clear that any recombinant was highly aggregated (FIG. 1A).

In addition, under non-reducing conditions, a rabbit anti-human IgG Fc polyclonal antibody as a primary antibody and a goat antibody as a secondary antibody against a commercial full-length DcR3-Fc (Abcam) produced using HEK293 cells as a host cell. When immunoblotting was performed using an anti-rabbit IgG antibody (Dako), most of them were present as aggregates as in the above-described purified product (Fig. 1B).

On the other hand, in the commercially available DcR3 FL-Fc (R&D) expressed in insect cells Sf21, almost no aggregation was observed, and a difference in aggregation was confirmed depending on the type of host cell producing wild-type DcR3 (FIG. 1C). R218Q-Fc having a R218Q mutation in which the 218th Arg of DcR3 FL-Fc (SEQ ID NO: 100) or human DcR3 (SEQ ID NO: 2) was substituted with Gln using insect cell S2 as a host cell by the following method ( US patents US6835814B1, US6965012B1, SEQ ID NO: 340), and S195-Fc (SEQ ID NO: 102) lacking HBD were prepared, and aggregation was evaluated. Any of the DNA fragment of human DcR3 (SEQ ID NO: 3), the DNA fragment in which the R218Q mutation was introduced into human DcR3 (SEQ ID NO: 111), or the DNA fragment of human DcR3 excluding HBD (SEQ ID NO: 107), and the linker sequence IEGRMD DNA The fragment (SEQ ID NO: 105) and the DNA fragment (SEQ ID NO: 71) of Fc (g1S) were artificially synthesized, pMTBiPV5-HisA (Thermo Scientific), Drosophila Expression System (Thermo Scientific), DcR3 FL-Fc, An S2 cell line stably expressing R218Q-Fc and S195-Fc was obtained. From the culture supernatant of the stable expression strain, affinity purification was performed by MabSelect SuRe in the same manner as above.

As a result, DcR3 FL-Fc, R218Q-Fc and S195-Fc produced from S2 cells hardly aggregated ( FIG. 1C ). Quantitative evaluation of aggregate content was performed by gel filtration chromatography (SEC) (TSKgel G3000 SWXL 7.8 mm x 300 mm) using HPLC (Shimadzu Corporation) (Tosoh Corporation). Table 1 shows the ratios (%) of monomers, aggregates, and decomposition products calculated from the peak area by analyzing S195-Fc derived from Expi293 and S2.

From the above results, it was found that when wild-type DcR3 was expressed in mammalian cells, the amount of aggregate production increased.

[Example 2] Preparation of non-aggregating DcR3 variants in mammalian cell expression system

Human DcR3 variants that do not aggregate in mammalian cell expression systems are not known so far. Therefore, while maintaining the activity of DcR3, the production of a DcR3 variant in which the amount of aggregates is reduced was attempted. DcR3 is a soluble molecule containing 300 residues, a signal peptide on the N-terminal side, followed by four cysteine rich domains characteristic of the TNF receptor superfamily (TNFRSF) (CRD1, CRD2, CRD3, CRD4) , and the C-terminal side includes a heparan sulfate binding domain (HBD) rich in basic amino acids including a heparan sulfate binding motif (FIG. 2, FIG. 3A). DcR3 ligands LIGHT, TL1A, and FasL all bind through CRD2 and CRD3 of DcR3. Therefore, while maintaining the region containing CRD2 and CRD3 of DcR3, CRD1 and / or CRD4 of DcR3 was substituted with a soluble decoy receptor osteoprotegerin (OPG), which is a flexible TNFRSF molecule, to prepare a DcR3 variant .

In human DcR3, CRD1 and CRD4 are amino acid sequences of human OPG, CRD2 and CRD3 are amino acid sequences of human DcR3, and chimera A (SEQ ID NO: 54) from which the amino acid sequence of HBD has been removed, and Fc sequence (IEGRMD g1S (IEGRMD g1S ( SEQ ID NO: 339) or g4PEK (SEQ ID NO: 74) in which the 228th Ser of the human IgG4 heavy chain represented by the EU index is substituted with Pro, the 235th Leu is substituted with Glu, and the 409th Arg is substituted with Lys chimeric A-Fc (Fig. 3C, SEQ ID NO: 80 or 82) fused to the chimeric A-Fc (Fig. 3C, SEQ ID NO: 80 or 82), human DcR3, wherein CRD1 is the amino acid sequence of human OPG, and CRD2, CRD3 and CRD4 are the amino acid sequence of human DcR3, , Chimera B (SEQ ID NO: 50) from which the amino acid sequence of HBD has been removed, and chimeric B-Fc (FIG. 3G, SEQ ID NO: 76) fused with an Fc sequence (IEGRMD g1S), in human DcR3, CRD1, CRD2 and CRD3 Chimeric C (SEQ ID NO: 52) in which the amino acid sequence of human DcR3, CRD4 as the amino acid sequence of human OPG, and the amino acid sequence of HBD was removed, and the Fc sequence (IEGRMD g1S) were fused to chimeric C-Fc (H in FIG. 3 ) , SEQ ID NO: 78), 103-123OPG (SEQ ID NO: 56) in which the amino acid sequence containing the 18th to 36th portions and 2 amino acids on the C-terminal side of CRD3 of chimera A was substituted with human OPG (SEQ ID NO: 56), and an Fc sequence (g4PEK) fused 103-123OPG-Fc(g4PEK) (FIG. 3D, SEQ ID NO: 84), various DcR3 variants having various structures were prepared by the following method. In addition, the DcR3 variant in which CRD4 is derived from DcR3 has the TS sequence that is the 194th to 195th amino acid sequence of DcR3 (SEQ ID NO: 2) at the C terminus of CRD4, and the DcR3 variant in which CRD4 is OPG is derived from CRD4 At the C-terminus, the SGNSESTQK sequence, which is the 186th to 194th amino acid sequence of OPG (SEQ ID NO: 14), is added.

A DNA fragment encoding a signal peptide sequence, chimera A, chimera B, chimera C, or 103-123OPG, from which the HBD sequence was removed (chimera A: SEQ ID NO: 53, chimera B: SEQ ID NO: 49, chimeric C: SEQ ID NO: 51 , 103-123OPG: SEQ ID NO: 55) is artificially synthesized and linked to a DNA fragment (SEQ ID NO: 338, 73) encoding an Fc sequence (IEGRMD g1S or g4PEK), in the same manner as in Example 1, pCIpuro vector DNA fragments encoding various DcR3 variants (chimeric A-Fc: SEQ ID NO: 79 or 81, chimeric B-Fc: SEQ ID NO: 75, chimeric C-Fc: SEQ ID NO: 77, 103-123OPG-Fc: sequence A plasmid into which No. 83) was inserted was obtained. In the same manner as in Example 1, the obtained plasmid was introduced into any host cell of Freestyle CHO-S cells, Freestyle 293F cells, and Expi293 cells, proteins were transiently expressed, and affinity purification was performed from the culture supernatant by MabSelectSuRe.

The ratio (%) of the monomers, aggregates, and decomposition products of the various DcR3 variants prepared was measured by SEC-HPLC in the same manner as in Example 1, or by SEC (Waters) using ACQUITY UPLC Protein BEH SEC 4.6 mm × 150 mm. was calculated by (Table 2).

As a result, the aggregates of any of the chimeric A-Fc, chimeric B-Fc, chimeric C-Fc, and 103-123OPG-Fc variants of DcR3 were reduced compared to that of wild-type DcR3. The effect of reducing aggregates was highest in chimeric A-Fc, followed by chimeric B-Fc and chimeric C-Fc. Therefore, it was shown that the substitution of OPG-derived CRD1 and CRD4 each had an effect of reducing aggregation, and that substitution of both CRD1 and CRD4 in combination had an effect of further reducing aggregation.

As a result of subjecting chimeric A-Fc, chimeric B-Fc, and chimeric C-Fc (all IEGRMD g1S) to SDS-PAGE under non-reducing and reducing conditions of 100 mM DTT, even when produced using mammalian cells as hosts , Smear or ladder was hardly confirmed in chimera A-Fc and chimera B-Fc (FIG. 4).

SEC (TSKgel G3000 SWXL 7.8 mm × 300 mm) using HPLC (Prominence, Shimadzu Corporation) for chimeric A-Fc (IEGRMD g1S or g4PEK) and 103-123OPG-Fc (g4PEK) to calculate absolute molecular weight (Tosoh Corporation) was analyzed. 50mmol/L phosphate buffer (pH7.0, 500mmol/L NaCl) is used for the mobile phase, and after separation at a flow rate of 0.75mL/min, for each peak detected at a wavelength of 215nm, Multi Angle Light Scattering (MALS) ) (miniDAWN TREOS, Wyatt Technology) was used to calculate the absolute molecular weight (% uncertain) using the intensity of scattered light detected. A result is shown in Table 3.

Since the molecular weights of the dimers predicted from the amino acid sequences of chimeric A-Fc (IEGRMD g1S or g4PEK) and 103-123OPG-Fc are 92.0 kDa, 90.1 kDa, and 90.5 kDa, respectively, any DcR3 variant exists as a dimer. it has been confirmed that

[Example 3] Analysis of N-type sugar chain addition rate of chimeric A-Fc (g4PEK) and evaluation of influence on aggregation

In the chimeric A-Fc (g4PEK) (SEQ ID NO: 82), the Asn predicted from the amino acid sequence of the N-type sugar chain addition is 1 to 3 locations (N131, N144, N157) and the Fc region of OPG-derived CRD4. A location N260 exists. Therefore, the presence or absence of N-type sugar chain addition of the chimeric A-Fc (g4PEK) was evaluated by the following method. For sample preparation, the N-type sugar chain of the reduced and alkylated chimeric A-Fc (g4PEK) is cleaved by PNGaseF treatment, and then the protein is digested with trypsin, end proteinase Asp-N, and chymotrypsin proteases. did The resulting peptide mixture was dissolved in 5% (v/v) acetonitrile/0.1% formic acid and analyzed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). Using MAGIC2000 HPLC (Michrom Bioresources) and LTQ Orbitrap XL ion trap-Orbitrap hybrid mass spectrometer (Thermo Scientific) equipped with a C18 reversed-phase column (0.2 mm x 150 mm) (GL Sciences), 5-65% (v) The analysis was performed by gradient elution of /v) acetonitrile/0.1% formic acid. The obtained peptide fragment was identified by performing MASCOT analysis (Matrix Science) on the amino acid sequence of the chimeric A-Fc (g4PEK). The N-type sugar chain addition Asn was converted to Asp by PNGaseF treatment and the mass increased by 0.984 Da as an index. Peptides with shifted MS spectrum peaks and sugar chain addition sites included in the peptide were identified. As a result, it was found that an N-type sugar chain was added to any Asn residues of N131, N144, N157 and N260. Of these, in almost all peptide fragments N157 and N260 had added sugar chains, but for N131 and N144, both fragments with and without added sugar chains were detected.

Next, for the purpose of evaluating the effect on the cohesiveness of the N-type sugar chain added to the Asn (N131, N144, N157) at the three sites of CRD4 derived from OPG, two sites of N131 and N144 or 3 sites of N131, N144 and N157 An amino acid substitution was prepared for removing the sugar chain at the site. As the two sugar chain removal products, N131S/N144S-Fc(g4PEK) (SEQ ID NO: 86) in which N131 and N144 are substituted with Ser, respectively, or T133A/S146A-Fc (g4PEK) (SEQ ID NO: 86) in which T133 and S146 are substituted with Ala, respectively 88) was prepared. As the three sugar chain removal products, N131S/N144S/N157S-Fc(g4PEK) (SEQ ID NO: 90) in which N131, N144, and N157 are substituted with Ser, respectively, or T133A/S146A/T159A in which T133, S146 and T159 are substituted with Ala, respectively -Fc(g4PEK) (SEQ ID NO: 92) was constructed. A DNA fragment of the signal peptide sequence and a DNA fragment encoding each sugar chain removal product (SEQ ID NOs: 85, 87, 89, 91) were artificially synthesized, inserted into the pCIpuro vector in the same manner as in Example 1, and introduced into Expi293 cells introduced. After transient expression of each sugar chain-removed body, affinity purification by MabSelectSuRe was performed from the culture supernatant. Each prepared sugar chain-removed product was analyzed by SEC-UPLC (device: ACQUITY UPLC, column; ACQUITY UPLC Protein BEH SEC 200 Å, 1.7 μm, 4.6×150 mm) (Waters). Table 4 shows the ratios (%) of monomers, aggregates, and decomposition products calculated from the peak area.

The aggregate content of the two sugar chains removed was slightly increased for any amino acid substitution compared to the chimeric A-Fc (Table 2), and the three sugar chain removed product showed a further increase in aggregates for any amino acid substitution. Confirmed. From this, it was shown that among the three N-type sugar chains (N131, N144, N157) added to OPG-derived CRD4, the sugar chain added to N157 in particular contributed to the reduction of chimera A aggregates. On the other hand, even when all three N-type sugar chains were removed, the proportion of aggregates was lower than that of DcR3 FL-Fc or S195-Fc (Table 1). Therefore, it was found that the N-type sugar chain and the OPG sequence each had an effect of reducing the aggregate of wild-type DcR3, and that combining both had the effect of further reducing the aggregate.

[Example 4] Evaluation of physical properties of various DcR3 variants

In order to analyze the cause of the aggregation reduction effect of the various DcR3 variants prepared in Examples 2 and 3, the following physicochemical analysis was performed. Hydrophobic interaction chromatography (HIC) is an analysis method in which the elution time from the column increases as the hydrophobicity of the protein surface increases. Using a hydrophobic chromatography column (TSKgel Butyl-NPR 4.6 mm × 35 mm) (Tosoh Corporation), buffer A (2 mmol/L ammonium sulfate, 20 mmol/L Tris buffer, pH 7.0) and buffer B (20 mmol/L Tris buffer) , pH 7.0) as a mobile phase, and the elution time (minutes) detected at a wavelength of 215 nm after separation of an 8 μg sample at a flow rate of 0.5 mL/min is shown in Table 5.

Compared to the 1-amino acid variant R218Q-Fc of the full-length DcR3 constructed from insect cell S2, S195-Fc from S2, chimeric A-Fc from mammalian cells (IEGRMD g1S, g4PEK), 103-123OPG-Fc (g4PEK) and The elution time was shortened in both of the sugar chain-removed substances at two places, and it is thought that the physical properties were improved due to the decrease in hydrophobicity, and the aggregates were reduced. In addition, it was found that the ratio of aggregates and the hydrophobicity of proteins were correlated with the hydrophobicity equivalent to or greater than that of R218Q-Fc in chimeric C and the three sugar chain removal products, which had a slightly attenuated aggregate reducing effect.

Subsequently, the thermal stability of the protein was evaluated by the differential scanning fluorescence (DSF) method. In a 96-well white microplate (BioRad), 9.5 µg of various DcR3 variants and 1 µL of SYPRO Orange Protein Gel Stain (invitrogen) diluted 50-fold with water were mixed in a 20 µL system, followed by a C1000 thermal cycler (BioRad). The temperature was raised from 20 °C to 95 °C by 0.5 °C for 10 seconds. Fluorescence at each temperature was detected in the FRET channel, and a melting curve (FIG. 5) and melting temperature (Tm value) (°C) (Table 6) were calculated using CFX Manager software (BioRad).

Compared to the 1-amino acid mutant R218Q-Fc of the full-length DcR3 produced in insect cell S2, the Tm value in S2-derived S195-Fc increased significantly and the thermal stability was improved, suggesting that the HBD region contributes to the thermal instability. became In addition, in the mammalian cell-derived chimeric A-Fc (IEGRMD g1S), since the Tm value was higher than that of S195-Fc, the DcR3 variant in which a part of the CRD of DcR3 was substituted with a part of the CRD of the OPG had aggregation properties. It was found that not only the decrease, but also the thermal stability of the protein was improved (Table 6, Fig. 5).

[Example 5] Reactivity evaluation of human normal cells and CHO cells of the DcR3 variant having the heparan sulfate binding domain (HBD) removed

It has been reported that wild-type DcR3 and its mutant FLINT (R218Q mutant) exhibit very poor in vivo kinetics in mice and cynomolgus monkeys (Drug Metabolism and Disposition, 2003, 31: p.502-507). One of these factors is thought to depend on the direct binding of DcR3 to heparan sulfate proteoglycans on the cell membrane by the heparan sulfate binding domain (HBD) present in wild-type DcR3 (J. Immunol., 2006, 176). : 173-180).

Therefore, the reactivity of the HBD-removed DcR3 variant S195-Fc, the chimeric A-Fc (IEGRMD g1S, g4PEK), and the 103-123OPG-Fc (g4PEK) to human primary cells and CHO cells, which are production cells, was evaluated by flow cytometry. (FCM). DcR3 FL-Fc containing the HBD region as a positive control, K194-Fc as a negative control, and an anti-DNP antibody (g4PEK) were used. K194-Fc (amino acid sequence: SEQ ID NO: 152, DNA nucleotide sequence: SEQ ID NO: 151) is an IEGRMD linker (SEQ ID NO: 106) and It is a protein fused with the amino acid sequence of Fc (g1S) (SEQ ID NO: 72), transiently expressed using Expi293 cells by the method described in Example 1, and from the culture supernatant using Mabselect SuRe (GE Healthcare) Purified. Anti-DNP antibody (g4PEK) was prepared by Clin. The variable region of the anti-2,4-dinitrophenol (DNP) antibody described in Cancer Res., 2005, 11(8), p.3126-3135 was inserted into a vector encoding an Fc sequence (g4PEK), and CHO cells was introduced and expressed, and protein A purified antibody from the culture supernatant was used. As human primary cells, HUVECs (Lonza) and Male Human Hepatocytes (BioreclamationIVT) were used. Human primary cells were cultured using a collagen-coated adhesion plate (IWAKI) using a medium designated for each cell according to the attached document. CHO cells were cultured in suspension using EX-CELL 325 PF CHO Serum-Free Medium (Sigma-Aldrich).

HUVECs and hepatocytes were exfoliated using 0.02% EDTA solution and a cell scraper, and passed through a cell strainer (40 µm). After washing with a buffer for FCM (PBS containing 1% BSA, 1 mmol/L EDTA, 0.05% NaN ₃ ), the CHO cells aliquoted from the exfoliated HUVECs, hepatocytes, and suspension culture were suspended in a buffer for FCM.

Next, it was seeded in a 96-well U-bottom plate (Falcon) so that the concentration of 1×10 ⁵ cells/well was reached, and each prepared Fc fusion protein was added to a concentration of 10 μg/mL, followed by reaction on ice for 1 hour. After washing the cells with a buffer for FCM, LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Molecular Probes) and 0.1 μg/mL Goat F(ab') ₂ Anti-Human IgG R-phycoerythrin Conjugate (Southern Biotech) was suspended and stained on ice for 1 hour. When staining HUVECs and hepatocytes, Human FcR Blocking Reagent (Miltenyi Biotech) was added. After washing the cells with a buffer for FCM, the fluorescence intensity was analyzed with a flow cytometer FACS Fortessa (BD Biosciences).

As a result of analyzing the PE staining intensity for the LIVE/DEAD Fixable Aqua Dead Cell Stain Kit-negative live cell fraction, DcR3 FL-Fc(IgG1) (R&D) showed significant binding to any of HUVECs, hepatocytes and CHO cells. . On the other hand, none of the DcR3 variants from which HBD was removed did not bind (FIG. 6).

[Example 6] Evaluation of in vivo kinetics of DcR3 variant mice

S195-Fc and each DcR3 variant shown in Table 7 were subjected to a mouse kinetic test. For chimeric A-Fc (g4PEK) and 103-123OPG-Fc (g4PEK), each DcR3 variant was prepared by stable expression of CHO cells by the method shown below. A DNA fragment of the signal peptide sequence and a DNA fragment encoding each DcR3 variant (SEQ ID NOs: 81, 83) were artificially synthesized, and produced by the method described in International Publication No. 2012/081628 in the same manner as in Example 1. It was inserted into a recombinant vector, and the vector was introduced into CHO cells by electroporation. Culture and drug selection were carried out in the usual manner, and those in which the proportion of viable cells recovered to about 98% were used as stable expression strains. After culturing this stable expression strain in a medium such as EX-CELL 325 PF CHO Serum-Free Medium (Sigma-Aldrich) for a certain period, the culture supernatant was recovered, and each DcR3 variant was purified by the method described in Example 1. .

S195-Fc and 10 mg/kg of each DcR3 variant were administered i.v. After administration (n=2 or 3), blood was collected from the caudal vein at any time after administration, and the concentration of S195-Fc and each DcR3 variant in the blood was measured by the following method. A biotinylated monkey anti-human IgG polyclonal antibody was reacted with immobilized streptavidin beads, and S195-Fc and each DcR3 variant in the bound serum were detected by Alexa Fluor 647-labeled monkey anti-human IgG polyclonal antibody. did Measurement was performed using Gyrolab xP workstation (Gyros AB), and each dynamic parameter was calculated by the moment analysis method. As a standard material for preparing a calibration curve, the same as the test material administered to the animals was used. Transitions of blood concentrations of S195-Fc produced in S2 cells and chimeric A-Fc (IEGRMD g1S) produced in CHO-S cells are shown in FIG. 7 and Table 7 for S195-Fc and each DcR3 variant. , Table 8 shows the half-life (h) of the disappearance phase after single administration and AUC0-∞ (㎍*h/mL), which is the area under the concentration-time curve to infinity, respectively.

Compared with the wild-type DcR3 control, S195-Fc, the blood concentration trend of the DcR3 variant was significantly improved ( FIG. 7 ). Although the half-life was almost equal, AUC0-∞ was improved more than 10-fold (Table 8).

A single dose of 0.5 mg/kg of wild-type DcR3 and FLINT (R218Q mutant) to CD-1 mice i. v. When administered, the blood half-lives are 1.2 hours and 3.1 hours, respectively, and AUC0-∞ (μg*h/mL) are reported to be 0.48 and 0.36, respectively (Drug Metabolism and Disposition, 2003. 31: p.502- 507.). Assuming linearity and comparing the concentration trends multiplied by the dose ratio, the DcR3 mutant showed high exposure to wild-type DcR3 and FLINT.

[Example 7] Evaluation of binding activity to DcR3 ligand

(1) Preparation of DcR3 ligand

A soluble recombinant of human, cynomolgus monkey or mouse DcR3 ligand (LIGHT, TL1A, FasL) was constructed (amino acid sequence: SEQ ID NOs: 114, 116, 118, 120, 122, 124, 126, 128, 130, DNA nucleotide sequence of: SEQ ID NOs: 113, 115, 117, 119, 121, 123, 125, 127, 129).

In human, cynomolgus monkey or mouse soluble recombinant LIGHT, a His tag (His10) and a GS linker (GGGSGGGSGGGSIEGR) are added to the N-terminus, and the extracellular region of LIGHT (human LIGHT: Asp74-Val240 (SEQ ID NO: 132), cynomolgus monkey LIGHT: Asp74-Val240 (SEQ ID NO: 134), mouse LIGHT: Asp72-Val239 (SEQ ID NO: 136)) was used.

Human, cynomolgus monkey or mouse soluble recombinant TL1A has a His tag (His6) and a GS linker (GGGSGGGSGGGS) added to the N-terminus, and the extracellular region of TL1A (human TL1A: Leu72-Leu251 (SEQ ID NO: 138), cynomolgus monkey TL1A: Leu72-Leu251 (SEQ ID NO: 140), mouse TL1A: Ile94-Leu270 (SEQ ID NO: 142)) were used.

In human, cynomolgus monkey or mouse soluble recombinant FasL, a His tag (His6) is added to the N-terminus, and an extracellular region of FasL (human FasL: Pro134-Leu281 (SEQ ID NO: 144), cynomolgus monkey) is added downstream thereto. A sequence linking FasL: Pro133-Leu280 (SEQ ID NO: 146), mouse FasL: Pro132-Leu279 (SEQ ID NO: 148)) was used.

The DNA fragment of the signal peptide sequence and the DNA sequence of the tag-added soluble DcR3 ligand were artificially synthesized (GENEWIZ), and using the In-Fusion HD Cloning Kit (Clontech), pCI-Hygro2.01 vector (Promega) pCI was inserted downstream of the CMV promoter (with some modifications), and E. coli DH5α competent cells (TOYOBO) were transformed.

The obtained plasmid was introduced into Expi293 cells (Thermo Scientific) to transiently express the protein. The plasmid was introduced using ExpiFectamine 293 (Thermo Scientific), and after 3 days of culture, the culture supernatant was recovered.

Human FasL and cynomolgus monkey FasL were introduced into host cells of Freestyle CHO-S cells (Thermo Scientific) to transiently express proteins. The plasmid was introduced using Freestyle MAX Reagent (Thermo Scientific), and after 3 days of culture, the culture supernatant was recovered.

Protein purification from the culture supernatant of Expi293 cells was performed using Ni Sepharose Fast Flow resin and His Buffer Kit (both GE Healthcare). The culture supernatant was passed through a column filled with resin, washed with a washing buffer (60 mmol/L imidazole, 20 mmol/L sodium phosphate, 0.5 mol/L NaCl, pH 7.4), and then elution buffer (250 mmol/L imidazole). dazole, 20 mmol/L sodium phosphate, 0.5 mol/L NaCl, pH7.4).

Protein purification from the culture supernatant of Freestyle CHO-S cells was performed using Complete His-Tag Purification Resin (Roche) and His Buffer Kit (GE Healthcare). The culture supernatant was passed through a column filled with resin, washed with a washing buffer (2 mmol/L imidazole, 20 mmol/L sodium phosphate, 0.5 mol/L NaCl, pH 7.4), and then elution buffer (250 mmol/L imidazole). dazole, 20 mmol/L sodium phosphate, 0.5 mol/L NaCl, pH7.4).

Each eluted fraction was replaced with PBS using a NAP column (GE Healthcare) and sterilized by passing through a 0.22 μm filter. Purity of the obtained purified protein was confirmed by SDS-PAGE. Multimer formation was analyzed by SEC-UPLC (device: ACQUITY UPLC, column; ACQUITY UPLC Protein BEH SEC 200 Å, 1.7 μm, 4.6 × 150 mm) (Waters). As a result, in almost all recombinant-soluble DcR3 ligands, Although the peak of the molecular weight corresponding to a trimer was confirmed, the peak of a trimer was not detected only in mouse LIGHT, but was a monomer.

(2) Measurement of binding activity using BIAcore

For the various wild-type DcR3 controls and DcR3 variants shown in Table 9, binding activity to DcR3 ligands (LIGHT, TL1A, FasL) was analyzed by SPR method using BIAcore T-100 (GE Healthcare). As a buffer, HBS-EP+buffer was used.

10000RU of anti-human antibody was immobilized on Series S Sensor Chip CM5 using Human Antibody Capture Kit (GE Healthcare), and then various wild-type DcR3 and DcR3 variants were flowed at 10 μL/min for 30 seconds to capture. On the other hand, a buffer solution containing no protein was flowed into the reference flow cell. Thereafter, as an analyte, human, cynomolgus monkey or mouse DcR3 ligand diluted to 0.08-80 nmol/L was flowed at 10 μL/min for 2 minutes to monitor binding, and then dissociation was monitored by flowing a buffer solution for 3 minutes. Next, 3 mol/L magnesium chloride was flowed at 20 µL/min for 1 minute to perform a regeneration reaction. Using BIAcore T-100 evaluation software, 1:1 binding model, each DcR3 ligand as a monomer (human LIGHT monomer: 20.8 kDa, human TL1A monomer: 22.1 kDa, human FasL monomer: 17.7 kDa, cynomolgus monkey LIGHT monomer: 20.8 kDa, cynomolgus monkey TL1A monomer: 22.0 kDa, cynomolgus monkey FasL monomer: 17.7 kDa, mouse LIGHT monomer: 20.9 kDa, mouse TL1A monomer: 21.5 kDa, mouse FasL monomer: 17.7 kDa) , each kinetic constant (k _a , k _d , K _D ) was calculated (Tables 10 to 12). As a result of the measurement, it was confirmed that various wild-type DcR3 and DcR3 variants, except for mouse LIGHT, which could not produce a trimer, bind to each DcR3 ligand in humans, cynomolgus monkeys, and mice.

[Example 8] Evaluation of binding activity to OPG ligands (RANKL, TRAIL)

It is known that RANKL is involved in bone resorption by binding to RANK, and TRAIL is involved in cell death by binding to TRAIL receptor. Binding activity to RANKL and TRAIL was analyzed by ELISA using the various prepared human DcR3 recombinants. As a positive control for binding to the OPG ligand, the C-terminal region defect K194-Fc, RANK-Fc (Enzo Life Sciences) and TRAIL R1-Fc (R&D Systems) produced in Example 5 were used. Anti-DNP antibody (IgG1) [Clin. Cancer Res., 2005, 11(8), p.3126-3135], the variable region of the anti-2,4-dinitrophenol (DNP) antibody described in was inserted into a vector encoding an IgG1 Fc, and introduced into CHO cells. and expressed, and protein A purified antibody was used.

In a 96-well plate (MAXISORP NUNC-IMMUNO PLATE, Thermo Scientific), 50 µL/well of anti-human IgG1 (American Qualex) prepared at 10 µg/mL in PBS was dispensed at a time, and allowed to stand at 4°C overnight for adsorption. After removing the solidified solution, 1% Block Ace prepared by dissolving 1 g of Block Ace powder (DS Pharma Biomedical Co., Ltd.) in 100 mL of water was dispensed at 100 μL/well at a time, allowed to stand at room temperature for 1 hour, and blocked, 0.1% It was washed three times with PBS containing Tween (hereinafter referred to as PBST). Next, in plate No. 1, various human DcR3 recombinants or various DcR3 variants and RANK-Fc diluted to 1 µg/mL with 1% BSA-PBS were mixed with 1% BSA-PBS, and in plate No. 2, 1% BSA-PBS. Various wild-type DcR3 or various DcR3 mutants and TRAIL R1-Fc diluted to [mu]g/mL were dispensed at a time of 50 µL/well, and allowed to stand at room temperature for 1 hour.

After washing each plate 3 times with PBST, RANKL (Peprotech) diluted to 0.64--50000 pg/mL with 1% BSA-PBS was applied to plate No. 1, and 0.64- to plate No. 2 with 1% BSA-PBS. TRAIL (Peprotech) diluted to 50000 pg/mL was dispensed at a time of 50 µL/well, and the mixture was left at room temperature for 1 hour.

After washing the plate 3 times with PBST, plate No. 1 with biotinylated anti-RANKL antibody (Peprotech) diluted to 0.4 μg/mL with 1% BSA-PBS was applied to plate No. 2 with 1% BSA-PBS. A biotinylated anti-TRAIL antibody (R&D) diluted to 0.4 µg/mL was dispensed at a time of 50 µL/well, and the mixture was left at room temperature for 1 hour.

After each plate was washed 3 times with PBST, 50 µL/well of streptavidin-HRP (PIERCE) diluted 10,000-fold with 0.1% Block Ace was dispensed at a time and allowed to stand at room temperature for 1 hour.

After each plate was washed 3 times with PBST, 50 µL/well of TMB+Substrate Chromogen (Dako) was dispensed at a time and allowed to stand at room temperature for 1 minute. A 0.5 mol/L sulfuric acid solution was dispensed at a time of 50 µL/well, the color reaction was stopped, and the absorbance at a sample wavelength of 450 nm and a reference wavelength of 570 nm was measured using a plate reader.

As a result, K194-Fc bound to OPG ligands RANKL and TRAIL, and RANK-Fc and TRAIL R1-Fc showed binding to RANKL and TRAIL, respectively, but DcR3 FL-Fc (R&D), S195-Fc, chimera A-Fc (IEGRMD g1S) and chimeric B-Fc (IEGRMD g1S) did not bind to either ligand ( FIG. 8 ). Together with the results of Example 7, the DcR3 variant prepared by substituting a portion of the CRD of DcR3 with a portion of the CRD of OPG exhibits binding activity equivalent to that of wild-type DcR3 to the ligand of DcR3, but does not bind to the ligand of OPG it was confirmed

[Example 9] Measurement of neutralizing activity of DcR3 ligand

By the method shown below, the neutralizing activity against LIGHT, TL1A, FasL of various wild-type DcR3 controls and DcR3 variants was measured. DNP antibody was prepared by the method described in Examples 5 and 8.

(1) LIGHT neutralization activity measurement

Using the human colon cancer cell HT-29 cell line (ATCC No.: HTB-38), the neutralizing activity of various wild-type DcR3 and various DcR3 variants was measured using IL-8 production by LIGHT addition as an index. For cell culture and evaluation of neutralization activity, McCoy's 5A medium (Gibco) to which 10% FBS (Gibco) and penicillin/streptomycin (Nacalai Tesque) were added was used.

After seeding the HT-29 cell line at 2×10 ⁴ cells/well in a 96-well adhesive culture plate (Sumitomo Bakelite), various wild-type DcR3 or various DcR3 variants to a final concentration of 0.1, 1, or 10 μg/mL was added. Thereafter, human LIGHT (Gly66-Val240) (described in JP2013153749) with a FLAG tag (DYKDDDDK) added to the N-terminus was added to a final concentration of 0.1 μg/mL, and the total amount of the culture solution was prepared to 200 μL/well. After culturing at 37°C for 3 days in a 5% CO ₂ incubator, the culture supernatant was recovered, and the IL-8 concentration in the culture supernatant was measured using AlphaLISA IL-8 Immunoassay Research Kit (Perkin Elmer).

As a result of the measurement, it was confirmed that the IL-8 production amount was decreased in a concentration-dependent manner in various human DcR3 mutants, indicating LIGHT neutralizing activity (FIG. 9).

(2) TL1A neutralizing activity measurement

Using human T cells, the neutralizing activity of various wild-type DcR3 and various DcR3 variants was measured using IFN-γ production by TL1A addition as an index. X-VIVO15 medium (Lonza) was used for cell culture and evaluation of neutralization activity.

Frozen healthy PBMCs (AllCells) were thawed in a water bath at 37°C, and suspended in a medium containing DNaseI (STEMCELL) warmed to 37°C. After shaking at 37° C. at low speed for 2 hours, T cells were isolated using EasySep Human T cell Enrichment Kit (STEMCELL). The isolated T cells were seeded at 1×10 ⁵ cells/well in a 96-well floating culture plate (Sumitomo Bakelite), and then various wild-type DcR3 controls or various DcR3 pieces to a final concentration of 0.1, 1, 10 μg/mL. Variant was added. Thereafter, recombinant His10 human TL1A prepared in Example 6 was added so that the final concentration was 0.1 µg/mL. Human IL-12 (Miltenyi Biotech) at a final concentration of 2 ng/mL and Recombinant Human IL-18 (MBL) at a final concentration of 50 ng/mL were added to prepare a total amount of culture solution of 200 µL/well. After culturing at 37°C for 3 days in a 5% CO ₂ incubator, the culture supernatant was recovered, and the concentration of IFN-γ in the culture supernatant was measured using AlphaLISA IFN-γ Immunoassay Research Kit (Perkin Elmer).

As a result of the measurement, it was confirmed that the amount of IFN-γ production was decreased in a concentration-dependent manner in various DcR3 mutants, thereby exhibiting TL1A neutralizing activity ( FIG. 10 ).

(3) Measurement of FasL neutralizing activity

Using the T-cell leukemia Jurkat cell line (DSMZ No.: ACC 282), the neutralizing activity of various wild-type DcR3 controls and various DcR3 variants was measured using FasL addition-induced apoptosis as an index. For cell culture and evaluation of neutralization activity, RPMI1640 medium (Nacalai Tesque Co., Ltd.) to which 10% FBS and penicillin/streptomycin were added was used.

After the Jurkat cell line was seeded at 5×10 ⁴ cells/well in a 96-well floating culture plate, various wild-type DcR3 or DcR3 variants were added to give final concentrations of 0.01, 0.1, and 1 μg/mL. Thereafter, recombinant human His6 Fas Ligand (CST Japan) was added to give a final concentration of 0.1 µg/mL, and the total amount of the culture solution was adjusted to 100 µL/well. In a 5% CO ₂ incubator, after overnight culture at 37° C., CellTiter-Glo (Promega) was added to a Jurkat culture plate at 100 μL/well, and ATP production was performed using a luminometer (Veritas, Promega). The number of viable cells using as an index was measured.

As a result of the measurement, it was confirmed that, in the various DcR3 mutants, the amount of ATP production derived from living cells increased in a concentration-dependent manner, thereby exhibiting FasL neutralizing activity ( FIG. 11 ).

[Example 10] Ligand neutralizing activity and binding activity of FasL binding lowering variants

Based on the chimeric A-Fc (g4PEK), among the DcR3 ligands, one amino acid substitution having binding and neutralizing activity to TL1A and LIGHT but having reduced binding and neutralizing activity to FasL was constructed and evaluated. Variants in which arbitrary amino acids of CRD2 and CRD3 shown in Table 13 were substituted with Ala or amino acids other than Ala were prepared, and neutralizing activity against DcR3 ligand was measured in the same manner as in Example 9. As a result, chimeric A-E57K-Fc(g4PEK) in which the 57th Glu from the N-terminus of the chimeric A-Fc(g4PEK) was substituted with Lys or Leu (amino acid sequence: SEQ ID NO: 94, DNA base sequence: SEQ ID NO: 93 ), chimeric A-E57L-Fc(g4PEK) (amino acid sequence: SEQ ID NO: 96, DNA base sequence: SEQ ID NO: 95), and chimeric A-R60K-Fc(g4PEK) (amino acid in which Arg at position 60 was substituted with Lys) Sequence: SEQ ID NO: 98, DNA nucleotide sequence: SEQ ID NO: 97) showed a decrease in FasL-selective neutralizing activity (Table 13).

Chimeric A-E57K-Fc(g4PEK) (SEQ ID NO: 94), Chimeric A-E57L-Fc(g4PEK) (SEQ ID NO: 96), Chimeric A-R60K-Fc(g4PEK) (SEQ ID NO: 96) with reduced FasL-selective neutralizing activity 98), in the same manner as in Example 7, binding activity to the DcR3 ligand was measured. As a result, in all variants of chimeric A-E57K-Fc (g4PEK), chimeric A-E57L-Fc (g4PEK) and chimeric A-R60K-Fc (g4PEK), only FasL binding was significantly reduced, TL1A and LIGHT Binding to was maintained (Fig. 12a, Fig. 12b, Fig. 12c).

[Example 11] Evaluation of aggregation of commercially available wild-type DcR3 in mammalian cells

In the same manner as in Example 1, a commercial full-length DcR3-Fc prepared using HEK293 cells as a host cell (Abcam), and a commercially available full-length DcR3-Fc prepared using CHO cells as a host cell (AdipoGen) , Electrophoresis was performed under reducing and non-reducing conditions with respect to an Fc fusion of a DcR3 molecule lacking about half of the C-terminal side of a commercially available HBD prepared using HEK293 cells as a host cell (Enzo). The molecular weight of the monomer predicted from the degree of polarity under reducing conditions was about 50 kDa to 60 kDa, but the degree of mobility under non-reducing conditions was significantly larger than the expected dimer molecular weight, and in all commercial products, most of them existed as aggregates (Fig. 13).

[Example 12] Preparation of FasL binding lowering variants

Among the DcR3 ligands, Glu (E57) at the 57th position from the N-terminus of the chimeric A-Fc(g4PEK) (SEQ ID NO: 82) in order to select variants that bind to TL1A and LIGHT and have reduced FasL-selective binding properties One amino acid substitution (E57X; X is any amino acid other than Glu) was prepared by substituting amino acids other than Glu. In addition, for E57K, E57L, E57R, and E57V in which E57 was substituted with any of Lys, Leu, Arg, and Val, the amino acid at the 53rd position from the N-terminus of the chimeric A-Fc (g4PEK) (SEQ ID NO: 82) in the vicinity Any of Trp (W53), Asn (N54) at position 54, Tyr (Y55) at position 55, Leu (L56) at position 56, Arg (R58) at position 58, further specific amino acid Z (Z is Asp, Glu) , Asn, Gln, Pro, Thr, Gly any of) two amino acid substitutions (E57X_W53Z, E57X_N54Z, E57X_Y55Z, E57X_L56Z, E57X_R58Z; each two amino acid substitutions are given a variant number shown in Figure 14a) and Fc (g4PEK) ) was prepared as a fusion.

Among them, chimeric A-E57K (amino acid sequence: SEQ ID NO: 66, DNA nucleotide sequence: SEQ ID NO: 65), chimeric A-E57R (amino acid sequence: SEQ ID NO: 180, DNA nucleotide sequence: SEQ ID NO: 179), chimeric A- E57V (amino acid sequence: SEQ ID NO: 182, DNA nucleotide sequence: SEQ ID NO: 181), chimera A-E57K_R58D (variant number: 45-10, amino acid sequence: SEQ ID NO: 184, DNA nucleotide sequence: SEQ ID NO: 183), chimera For A-E57K_R58E (variant number: 45-18, amino acid sequence: SEQ ID NO: 186, DNA nucleotide sequence: SEQ ID NO: 185) and chimera A, fusions with some of the Fc sequences introduced with various mutations shown in Table 14 were prepared. (Base sequence: SEQ ID NOs: 213, 217, 219, 221, 223, 227, 231, 233, 235, 237, 255, 259, 261, 263, 265, 149, 167, 169, 171, 173, 175, 177 , amino acid sequence: SEQ ID NOs: 214, 218, 220, 222, 224, 228, 232, 234, 236, 238, 256, 260, 262, 264, 266, 317, 319, 320, 321, 322, 324, 326, 327, 328, 329, 331, 333, 334, 335, 336, 150, 168, 170, 172, 174, 176, 178). In Table 14, E216 represents the 216th Glu of a human IgG1 heavy chain represented by the EU index. C220S, M252Y, S254T, T256E, N434A, L234A, L235A, and G237A of the mutation introduction site, respectively, of the human IgG1 heavy chain represented by the EU index, Cys at position 220 of the Ser, Met at position 252 as Tyr, and at position 254 It indicates that Ser is replaced with Thr, that Thr at position 256 is replaced with Glu, Asn at position 434 is replaced with Ala, Leu at position 234 with Ala, Leu at position 235 with Ala, and Gly at position 237 with Ala.

Chimeric A-Fc (g1S) (base sequence: SEQ ID NO: 149, amino acid sequence: SEQ ID NO: 150) is a chimeric A-Fc (IEGRMD g1S) comprising the linker sequence IEGRMD (SEQ ID NO: 106) prepared in Example 2 (SEQ ID NO: 80) to which the IEGRMD sequence was deleted was prepared by stable expression of CHO cells by the method described in Example 6. Chimera A-Fc(Eg1S) (base sequence: SEQ ID NO: 167, amino acid sequence: SEQ ID NO: 168) was obtained by linking chimera A (SEQ ID NO: 54) and Eg1S to the stability of CHO cells by the method described in Example 6 It was produced by expression.

Chimeric A-Fc (Eg1S YTE, Eg1S N434A, Eg1S LALAGA, Eg1S LALAGANA) (base sequence: SEQ ID NO: 169, 171, 175, 177, amino acid sequence: SEQ ID NO: 170, 172, 176, 178) is, A sequence obtained by fusion of each Fc to the C terminus of No. 54) was obtained by transient expression in CHO-S cells.

Chimeric A-Fc (g1S YTE, g1S N434A, g1S LALAGA, g1S LALAGANA) (amino acid sequence: SEQ ID NO: 314, 315, 174, 316) was obtained by fusion of each Fc to the C terminus of chimeric A (SEQ ID NO: 54), respectively. Sequences were obtained by transient expression of Expi293 cells.

A plasmid expressing a fusion of each amino acid substitution and Fc (g4PEK) is prepared using either DNA sequence of chimeric A-Fc (g4PEK) or E57X-Fc (g4PEK; X is K, R or V) as a template, and the mutation site is Using PCR primers designed to contain, PCR amplification of two regions from the NheI site to the site into which the amino acid substitution was introduced and from the site to the site into which the amino acid substitution was introduced to the SalI site was PCR-amplified, respectively, in the same manner as in Example 1. It was constructed by inserting it under the CMV promoter of the pCIpuro vector.

A fusion of each amino acid substitution and each Fc of Eg1S YTE, Eg1S N434A, or Eg1S LALAGANA is a PCR primer designed to contain a mutation site using any DNA sequence of chimeric A-Fc having each mutated Fc sequence as a template. Using PCR amplification of the two regions from the EcoRI site to the site into which the amino acid substitution was introduced, and from the site to the site where the amino acid substitution was introduced to the Bsu36I site, respectively, as a template, EcoRI and Bsu36I sites of a chimeric A-variety Fc vector were used. was inserted to construct a plasmid.

The fusion of each amino acid substitution and each Fc of g1S YTE, g1S N434A, or g1S LALAGANA uses any of the above-described amino acid substitution-Fc DNA sequences as a template, and using PCR primers designed to exclude E216, EcoRI site A plasmid was prepared by PCR-amplifying two regions from to the Glu site to be deleted and from the Glu site to the Bsu36I site to be deleted, respectively, and inserted into the EcoRI and Bsu36I sites of each amino acid substitution-Fc vector used as a template.

Each plasmid was introduced into Expi293 cells for transient expression, and affinity purification was performed from the culture supernatant by MabSelect SuRe. The content of each of the monomers, aggregates, and degradation products of the prepared chimeric A-Fc, each 1 or 2 amino acid substitution-Fc was measured by SEC-UPLC (ACQUITY UPLC Protein BEH SEC 4.6 mm × 150 mm) (Waters) in the same manner as in Example 2. Co.) or SEC-HPLC (TSKgel SuperSW3000 4.0 µm, 4.6 mm × 300 mm) (Tosoh Corporation) was calculated from the analyzed peak area.

As a result, most of the prepared chimeric A-Fc and 1 and 2 amino acid substitution-Fc having various mutated Fc maintained a lower aggregate content than S195-Fc ( FIGS. 14A, 14B and 14C ).

[Example 13] Evaluation of binding activity to DcR3 soluble trimeric ligand

DcR3-Fc, S195-Fc, soluble human LIGHT trimer, soluble human TL1A trimer and soluble type for various chimeric A-Fc having different Fc sequences and various FasL binding lowering variants prepared in Example 12 The evaluation of binding activity to human FasL trimer was carried out with some modifications of the method described in Example 7.

(1) Preparation of DcR3 Soluble Trimeric Ligand

For human soluble recombinant LIGHT, a sequence (FLAG-LIGHT) in which a FLAG tag (DYKDDDDK) was added to the N-terminus, and the extracellular region of LIGHT (Asp74-Val240) (SEQ ID NO: 132) was linked downstream thereof (FLAG-LIGHT) was used (Base sequence: SEQ ID NO: 305, amino acid sequence: SEQ ID NO: 306). Human soluble recombinant TL1A is a sequence (His6) in which a His tag (His6) and a GS linker (GGGSGGGSGGGS) are added to the N-terminus, and the TL1A extracellular region (Leu72-Leu251) (SEQ ID NO: 138) is ligated downstream thereof. -TL1A) was used (base sequence: SEQ ID NO: 115, amino acid sequence: SEQ ID NO: 116). Each plasmid was prepared in the same manner as in Example 7, and transient expression by Expi293 cells was performed.

FLAG-LIGHT was purified using ANTI-FLAG M2 Affinity Gel (Sigma). The culture supernatant was passed through a column filled with resin, washed with a washing buffer (50 mM Tris HCl, 150 mM NaCl, pH 7.4), and eluted with an eluent (0.1 M glycine hydrochloride, pH 3.5).

His6-TL1A was purified by the following method. The culture supernatant was passed through a column filled with Complete His-Tag Purification Resin (Roche), washed with a washing buffer (50mM NaH2PO4 pH8.0, 300mM NaCl), and then the eluate (50mM NaH2PO4 pH8.0, 300mM NaCl, 250mM) imidazole). Using a NAP column (GE Healthcare), the eluate substituted with PBS was passed through a column filled with Ni Sepharose Fast Flow resin (GE Healthcare), and washing prepared using a His Buffer Kit (GE Healthcare) was used. After washing with an elution buffer (60 mM imidazole, 20 mM sodium phosphate, 0.5 M NaCl, pH 7.4), it was eluted with an elution buffer (250 mM imidazole, 20 mM sodium phosphate, 0.5 M NaCl, pH 7.4).

Each eluted fraction of FLAG-LIGHT and His6-TL1A was replaced with PBS using a NAP column (GE Healthcare) and sterilized by passing through a 0.22 μm filter. The obtained purified protein was subjected to gel filtration chromatography (SEC) (TSKgel G3000 SWXL 7.8 mm × 300 mm) (Tosoh Corporation) using HPLC (Shimadzu Corporation) to fractionate the trimer fraction.

Human soluble recombinant FasL is human His6 Fas Ligand/TNFSF6 (Cell Signaling TECHNOLOGY) in which a His tag (His6) is added to the N-terminus, and an extracellular region of FasL (Pro134-Leu281) (SEQ ID NO: 144) is linked thereto. g) was used. SEC-MALS was performed by the method similar to Example 2, and it confirmed that it was a trimer.

(2) Measurement of binding activity using BIAcore

DcR3-Fc (R&D Corporation) and S195-Fc were evaluated as described in Example 7. Chimeric A-g4PEK (SEQ ID NO: 82) was constructed in Example 6. Chimeric A-Fc (g1S, Eg1S) was prepared in Example 12. As various FasL binding lowering variants, the Fc (g4PEK) fusions prepared in Example 9 or Example 12 were evaluated.

The binding activity to the human DcR3 trimer ligand was analyzed by the SPR method. BIAcore T-100 (GE Healthcare) for measuring binding activity to human LIGHT and human TL1A, BIAcore T-100 (GE Healthcare) or BIAcore T-200 (GE Healthcare) for measuring binding activity to human FasL g) were used respectively. HBS-EP+buffer was used as the buffer.

10000RU of anti-human antibody was immobilized on the Series S Sensor Chip CM5 using the Human Antibody Capture Kit (all GE Healthcare), and then various wild-type DcR3 and DcR3 variants were flowed at 10 μL/min for 30 seconds to capture. On the other hand, a buffer solution containing no protein was flowed into the reference flow cell. Thereafter, as an analyte, each human DcR3 trimer ligand diluted to 0.02-80 nmol/L was flowed for 2 minutes at 30 µL/min to monitor binding, and then, a buffer solution was flowed for 3 minutes to monitor dissociation. Next, 3 mol/L magnesium chloride was flowed at 30 µL/min for 1 minute to perform a regeneration reaction. BIAcore T-100 evaluation software, a 1:1 binding model was used to measure binding activity to human LIGHT and human TL1A, and the ligand was a trimer (human LIGHT trimer: 62.4 kDa, human TL1A trimer: 66.2 kDa). , each kinetic constant (k _a , k _d , K _D ) was calculated. BIAcore T-100 evaluation software, 1:1 binding model or BIAcore T-200 evaluation software, 1:1 binding model was used to measure binding activity to human FasL, and the ligand was used as a monomer (human FasL monomer: 19.8 kDa). , each kinetic constant (k _a , k _d , K _D ) was calculated.

As a result, it was confirmed that chimeric A-Fc (g1S), chimeric A-Fc (Eg1S), and chimeric A-Fc (g4PEK) all bind to each DcR3 trimer ligand ( FIG. 15A ).

Among the FasL binding lowering variants (g4PEK) prepared in Example 12, one amino acid substitution chimera A-E57K-Fc (g4PEK) (amino acid sequence: SEQ ID NO: 94, DNA nucleotide sequence: SEQ ID NO: 93) (hereinafter “E57K”) -Fc"), chimeric A-E57L-Fc(g4PEK) (amino acid sequence: SEQ ID NO: 96, DNA base sequence: SEQ ID NO: 95) (hereinafter referred to as "E57L-Fc" in some cases); Chimeric A-E57R-Fc (g4PEK) (amino acid sequence: SEQ ID NO: 190, DNA base sequence: SEQ ID NO: 189) (hereinafter sometimes referred to as "E57R-Fc"), chimeric A-E57V-Fc (g4PEK) ( Amino acid sequence: SEQ ID NO: 192, DNA nucleotide sequence: SEQ ID NO: 191) (hereinafter sometimes referred to as "E57V-Fc"), chimeric A-E57A-Fc(g4PEK) (amino acid sequence: SEQ ID NO: 288, DNA base Sequence: SEQ ID NO: 287) (hereinafter sometimes referred to as "E57A-Fc"), chimeric A-E57F-Fc (g4PEK) (amino acid sequence: SEQ ID NO: 290, DNA base sequence: SEQ ID NO: 289) (hereinafter "E57F") -Fc"), chimeric A-E57H-Fc(g4PEK) (amino acid sequence: SEQ ID NO: 292, DNA base sequence: SEQ ID NO: 291) (hereinafter referred to as "E57H-Fc" in some cases); Chimeric A-E57I-Fc (g4PEK) (amino acid sequence: SEQ ID NO: 294, DNA nucleotide sequence: SEQ ID NO: 293) (hereinafter sometimes referred to as "E57I-Fc"), chimeric A-E57M-Fc (g4PEK) ( Amino acid sequence: SEQ ID NO: 296, DNA nucleotide sequence: SEQ ID NO: 295) (hereinafter sometimes referred to as "E57M-Fc") and 2-amino acid substitution chimera A-E57K_R58D-Fc(g4PEK) (variant number: 45-10 , amino acid sequence: SEQ ID NO: 194, DNA nucleotide sequence: SEQ ID NO: 193) (hereinafter sometimes referred to as "45-10-Fc"), chimera A-E57K_ R58T-Fc (g4PEK) (variant number: 45-11, amino acid sequence: SEQ ID NO: 298, DNA base sequence: SEQ ID NO: 297) (hereinafter sometimes referred to as “45-11-Fc”), chimeric A- E57K_R58E-Fc (g4PEK) (variant number: 45-18, amino acid sequence: SEQ ID NO: 196, DNA base sequence: SEQ ID NO: 195) (hereinafter sometimes referred to as “45-18-Fc”), chimeric A- E57L_R58E-Fc(g4PEK) (variant number: 46-4, amino acid sequence: SEQ ID NO: 300, DNA base sequence: SEQ ID NO: 299) (hereinafter sometimes referred to as “46-4-Fc”), chimeric A- E57R_R58D-Fc (g4PEK) (variant number: 82-5, amino acid sequence: SEQ ID NO: 198, DNA base sequence: SEQ ID NO: 197) (hereinafter sometimes referred to as “82-5-Fc”), chimeric A- E57V_R58T-Fc (g4PEK) (variant number: 85-6, amino acid sequence: SEQ ID NO: 302, DNA base sequence: SEQ ID NO: 301) (hereinafter sometimes referred to as “85-6-Fc”), chimeric A- E57V_R58E-Fc (g4PEK) (variant number: 85-8, amino acid sequence: SEQ ID NO: 304, DNA base sequence: SEQ ID NO: 303) (hereinafter sometimes referred to as “85-8-Fc”) is chimeric Compared with A-Fc (g4PEK), the K _D value for LIGHT trimer and TL1A trimer is less than 3 times, and the K _D value for FasL trimer is more than 3 times or the Rmax value is lowered to less than 5 and FasL-selective binding was confirmed to be lowered (Fig. 16a, Fig. 16b).

Among them, E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc, 45-10-Fc, 45-18-Fc, 82-5-Fc (all Fc is g4PEK) were analyzed by SEC-HPLC (column; TSKgel). G3000 SWXL 7.8 mm x 300 mm, Tosoh Corporation, each kinetic constant when purified to 95% or more of monomers by HPLC (Shimazu Seisakusho) and used for BIAcore measurement is shown in Fig. 15B.

[Example 14] Evaluation of neutralizing activity of DcR3 soluble ligand

Neutralizing activity evaluation for soluble human LIGHT, soluble human TL1A, and soluble human FasL of various DcR3 variants was performed by partially changing the method described in Example 9.

(1) Construction of various chimeric A-Fc having different Fc sequences

The chimeric A-Fc (g1S, Eg1S) prepared in Example 12 was used. Each chimeric A-Fc prepared in Example 12 by fusing each Fc of Eg1S YTE, Eg1S N434A, Eg1S LALAGA, or Eg1S LALAGANA shown in Table 14 on the C-terminal side of chimera A, respectively, is a CHO-S cell transient What was produced by expression and purified to 95% or more of monomers by SEC (Superdex 200 Increase 10/300 GL) (GE Healthcare) using AKTApurifier (GE Healthcare) was used.

(2) Soluble human LIGHT neutralizing activity

For chimeric A-Fc (g1S, Eg1S, Eg1S YTE, Eg1S N434A, Eg1S LALAGA, Eg1S LALAGANA, g4PEK), the inhibitory activity on LIGHT-dependent CXCL10 production from IFN-γ-stimulated long myofibroblasts (Lonza) was evaluated. Long myofibroblasts were cultured in a collagen I coat flask (BD) using the SmGM-2 Bullet Kit (Lonza) medium. After seeding the cells at 1×10 ⁴ cells/well in a collagen I coat 96-well plate (BD), the final concentration of IFN-γ was 10 ng/mL, and the final concentration of the trimeric FLAG-LIGHT prepared in Example 13. 20ng/mL, each final concentration of each of the various DcR3 variants is 19.5, 78.1, 313, 1250, 5000, 20000ng/mL, or 4.88, 19.5, 78.1, 313, 1250, 5000ng/mL. Incubated for one day. The culture supernatant was recovered, and the CXCL10 concentration in the culture supernatant was measured using the CXCL10/IP-10 (human) AlphaLisa Detection Kit (Perkin Elmer). As a result, it was confirmed that the chimeric A-Fc having any Fc sequence also inhibited CXCL10 production in a concentration-dependent manner and had neutralizing activity against soluble LIGHT ( FIGS. 17A and 17B ).

For FasL binding lowering variants E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc (all Fc is g4PEK), in the same manner as in Example 9, LIGHT-dependent IL-8 from HT-29 cells Inhibitory activity on production was evaluated. As a result, it was confirmed that any of the variants had a neutralizing activity against soluble LIGHT by inhibiting IL-8 production in a concentration-dependent manner (FIG. 17c).

For FasL binding lowering variants 45-10-Fc, 45-18-Fc, 82-5-Fc (all Fc is g4PEK), like chimeric A-Fc, LIGHT-dependent from IFN-γ-stimulated long myofibroblasts The inhibitory activity on CXCL10 production was evaluated under the conditions of 2×10 ⁴ long myofibroblasts per well and 19.5, 78.1, 313, 1250, 5000, and 20000 ng/mL of each final concentration of the test substance. As a result, it was confirmed that, for any of the variants, CXCL10 production was inhibited in a concentration-dependent manner, thereby having neutralizing activity against soluble LIGHT (FIG. 17d).

(3) Soluble human TL1A neutralizing activity

In the same manner as in Example 9, the inhibitory activity of various DcR3 variants on TL1A-dependent IFN-γ production from IL-12 and IL-18-stimulated human T cells was evaluated. As a result, it was confirmed that the chimeric A-Fc having any Fc sequence also inhibited IFN-γ production in a concentration-dependent manner and had neutralizing activity against soluble TL1A ( FIGS. 18A and 18B ). In addition, with respect to FasL binding lowering variants E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc, 45-10-Fc, 45-18-Fc, 82-5-Fc (all Fc is g4PEK), Similarly, it was confirmed to have a neutralizing activity against soluble TL1A ( FIGS. 18c and 18d ).

(4) Soluble human FasL neutralizing activity

In the same manner as in Example 9, the inhibitory activity of various DcR3 variants against FasL-dependent apoptosis of Jurkat cells or Jurkat subclone A3 cells was evaluated. As a result, it was confirmed that the chimeric A-Fc having any Fc sequence also inhibited the cell death of A3 cells in a concentration-dependent manner, and had neutralizing activity against soluble FasL ( FIGS. 19A and 19B ). On the other hand, for FasL binding lowering variants E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc, 45-10-Fc, 45-18-Fc, 82-5-Fc (all Fc is g4PEK), In all of them, the inhibitory activity against Jurkat cell death was remarkably reduced, and it was confirmed that the neutralizing activity against soluble FasL was selectively decreased (Fig. 19c, D).

[Example 15] Evaluation of binding activity to DcR3 membrane ligand

The binding activities of various DcR3 variants to membrane-type human LIGHT, membrane-type human TL1A, and membrane-type human FasL were evaluated by flow cytometry using a forced expression strain of membrane-type ligand. For membranous human LIGHT forced expression HEK293 cells, those described in US8974787 were used. Membrane-type TL1A and membrane-type FasL were PCR amplified by adding a Met residue and a FLAG tag (DYKDDDDK) to the N-terminal end of each commercially available ORF clone (Origene) to a template, and PCR amplification (base sequence: SEQ ID NOs: 307, 309, amino acid sequence: SEQ ID NOs: 308 and 310), were inserted downstream of the CMV promoter of the pCIpuro vector using the In-Fusion HD Cloning Kit (Clontech), and E. coli DH5α competent cells (TOYOBO) were transformed.

The obtained plasmid was introduced into CHO-K1 cells (ECACC) using Nucleofector and Cell Line Nucleofector Kit T (all manufactured by Lonza), and subjected to drug selection with 10 μg/mL puromycin (Thermo Fisher Scientific). The obtained drug-resistant cells were stained with DyLight 488-labeled anti-human TL1A antibody (Novus Biologics) or APC-labeled anti-human FasL antibody (BD Pharmingen), and high-expression fractions were sorted using a cell sorter (Sony). After expansion culture, the cells were stained with PE-labeled anti-human TL1A antibody (Novus Biologics) or PE-labeled anti-human FasL antibody (BioLegend), followed by sorting again, to obtain a cell line high expressing membranous human TL1A or membranous human FasL.

The binding activity of various DcR3 variants to the membranous ligand forced expression strain and host cell was evaluated in the same manner as in Example 5, changing the following conditions. HEK293 and membrane-type LIGHT forced expression strains were reacted with 1 μg/mL of each protein and 10 ng/mL of secondary antibody Goat F(ab') ₂ Anti-Human IgG R-phycoerythrin Conjugate (Southern Biotech) at 10 ng/mL. For the membrane-type TL1A and membrane-type FasL forced expression strains, each protein was reacted at 1 or 10 µg/mL and the secondary antibody was reacted at 0.1 or 1 µg/mL, respectively.

As a result, all of the chimeric A-Fc (g1S, Eg1S, g4PEK) did not react with 293 or CHO-K1 cells, which are host cells, but reacted specifically with each membrane ligand forcible expression strain. Accordingly, it was confirmed that all of the various chimeras A having different Fc sequences had binding activity to the membrane-type DcR3 ligand ( FIGS. 20A and 20B ). In addition, ligands for FasL binding lowering variants E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc, 45-10-Fc, 45-18-Fc, 82-5-Fc (all Fc are g4PEK) As a result of similarly evaluating the binding activity to membranous human LIGHT and membranous human TL1A, it was confirmed that while the binding activity was maintained, the binding activity to membranous human FasL was significantly reduced except for E57L ( 21a, 21b, 21c).

[Example 16] Evaluation of binding activity to DcR3 primary ligand

The binding activity of the various DcR3 variants to each DcR3 ligand derived from the primary cell was evaluated by the following method.

(1) Primary LIGHT binding activity

It is known that the expression of membranous LIGHT is induced in activated human T cells (The Journal of Immunology, 2004, 173: p.502-507.). Frozen healthy PBMCs (AllCells) were thawed and stimulated overnight with PMA (Sigma) at a final concentration of 50 ng/mL and ionomycin (Sigma) at a final concentration of 1 µg/mL, followed by expression in CD3-positive T cells. The binding of the chimeric A-Fc(Eg1S) to the induced membranous LIGHT was evaluated by the following method. As a negative control, the anti-DNP antibody (IgG1) described in Example 8 was used. The stimulated PBMCs were recovered, reacted with Human FcR Blocking Reagent (Miltenyi Biotech), BV421 labeled CD3 antibody (BD Pharmingen), 7-AAD Staining Solution (BD Pharmingen), and Alexa Fluor488 Antibody Labeling Kit (Thermo Scienific) ) at a final concentration of 0.4 μg/mL of chimeric A-Fc or anti-DNP antibody, respectively, was added. After the reaction, the cells were washed, and the fluorescence intensity of Alexa Fluor488 in the CD3-positive T cells of the live cell fraction was analyzed with a flow cytometer. To confirm the expression of membrane LIGHT in induction-stimulated PBMCs, after reacting the induction-stimulated PBMCs with Human FcR Blocking Reagent (Miltenyi Biotech), BV510-labeled CD3 antibody (BioLegend), 7-AAD Staining Solution (BD) Pharmingen) and PE-labeled LIGHT antibody (LSBio) or PE-labeled mouse IgG1κ isotype control antibody (BioLegend) were added, respectively, followed by reaction, followed by washing and PE in the CD3-positive T cells of the viable cell fraction with a flow cytometer. fluorescence intensity was analyzed.

As a result, it was confirmed that the expression of LIGHT was induced on CD3-positive human T cells of the induction-stimulated PBMC, and it was confirmed that the Alexa Fluor488-labeled chimera A-Fc was bound to the membranous LIGHT on the activated CD3-positive human T cells. (A, B in Fig. 22).

(2) primary TL1A binding activity

HUVEC cells (Lonza) cultured in the same manner as in Example 5 were treated with Recombinant Human IL-1 alpha (R&D) at a final concentration of 10 ng/mL, and TACE inhibitor TAPI-1 (Calbiochem) at a final concentration of 20 µM. Binding of the chimeric A-Fc (IEGRMD g1S) to the membrane-type TL1A induced by treatment for 24 hours was evaluated by a competition experiment with the TL1A antibody 1D1 1.31 (US2015/0132311). TL1A antibody 1D1 1.31 was transiently expressed using Expi293 cells by the method described in Example 1 by linking the amino acid sequences of VL and VH described in US2015/0132311 to the constant region of human IgG1, and Mabselect SuRe from the culture supernatant (GE Healthcare, Inc.) was used as purified. As a negative control, the anti-DNP antibody (IgG1) described in Example 8 was used. The recovered cells were reacted with Human FcR Blocking Reagent (Miltenyi Biotech) and labeled with Zenon Alexa Fluor 647 Human IgG Labeling Kit (Molecular Probes), anti-DNP antibody, chimeric A-Fc, TL1A antibody at the final concentration It was added so as to be 8.3 μg/mL. After the reaction, washing was performed, and the fluorescence intensity of Alexa Fluor 647 was analyzed with a flow cytometer. In the competitive conditions, any of the unlabeled DNP antibody, the TL1A antibody, and the chimeric A-Fc was reacted beforehand with a final concentration of 50 µg/mL.

As a result, the binding of labeled chimeric A-Fc competes with the unlabeled TL1A antibody, and the binding of the labeled TL1A antibody competes with the unlabeled chimeric A-Fc. It was confirmed that there is (FIG. 23).

(3) primary FasL binding activity

Binding of the chimeric A-Fc (IEGRMD g1S) to primary soluble FasL produced from human T cells induced by activation-induced cell death (AICD) was evaluated by the following method. Primary soluble FasL was adjusted by the following method. That is, in the same manner as in Example 9, human T cells isolated from frozen healthy PBMCs were seeded in 96-well U-bottom plates (BD) at 2×10 ⁴ cells/well, and PHA with a final concentration of 1 μg/mL After culturing in -L (eBioscience) for 24 hours, IL-2 (Peptrotech) at a final concentration of 1 μg/mL was added and cultured for 5 days. After 5 days, cells were harvested, and seeded in 96-well U-bottom plates immobilized with 5 µg/mL of anti-CD3 antibody OKT3 (BioLegend), AICD was induced and cultured overnight, the culture supernatant was recovered and sterilized water Using Amicon Ultra-15 (Millipore) with a molecular weight cutoff of 10 kDa, pre-rinsed, the solution volume was concentrated to 1/10.

After blocking a 96-well immune plate (Thermo Scientific) immobilized with 10 µg/mL anti-human IgG antibody (American Qualex) with 1% Block Ace (DS Pharma Biomedical), 20 µg/mL Chimera A -Fc (IEGRMD g1S) or Fas-Fc (R&D) was captured. After washing, the recombinant FasL (abcam) or 10-fold concentrated AICD culture supernatant used as a standard was reacted, and after washing, a biotinylated anti-FasL antibody (abcam) was reacted. After washing, react with streptavidin-HRP (PIERCE), wash again, add TMB solution (abcam) to develop color, stop the color reaction with 2N sulfuric acid solution, and measure the absorbance at 450 nm measured.

Fig. 24A shows the result of the plate encapsulating the chimeric A-Fc (IEGRMD g1S), and Fig. 24B shows the result of the plate encapsulating Fas-Fc. CD3 of 10×AICDsup. indicates that the culture supernatant of T cells induced with AICD was used under anti-CD3 antibody OKT3 stimulation conditions, and none indicates that the culture supernatant of T cells that did not induce AICD was used. For the plates in which either chimeric A-Fc or Fas-Fc were captured, FasL was detected only in the culture supernatant of AICD-induced T cells. From this point, it was confirmed that the chimeric A-Fc binds to soluble FasL produced from AICD-induced T cells ( FIGS. 24A and 24B ).

[Example 17] Evaluation of physical properties of DcR3 variants

Various chimeric A-Fc (g1S, Eg1S, Eg1S YTE, Eg1S N434A, Eg1S LALAGANA, Eg1S LALAGA, g4PEK) and FasL binding lowering variants E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc, 45-10 For -Fc, 45-18-Fc, 82-5-Fc (all g4PEK), in the same manner as in Example 4, calculation of elution time (min) by hydrophobic interaction chromatography (HIC) and differential scanning The Tm value (°C) was calculated by the fluorescence meter (DSF) method.

As a result, it was confirmed that there was no significant effect on hydrophobicity by introducing a plurality of amino acid mutations into the IgG1 Fc sequence. All of the elution times of the FasL binding lowering variants were not significantly different from those of the chimeric A-g4PEK, so it was confirmed that there was no effect on hydrophobicity by 1 or 2 amino acid substitutions (FIG. 25).

Regarding the Tm value by DSF, the effect on thermostability by the introduction of YTE mutation during amino acid substitution or insertion of the IgG1 Fc sequence was confirmed. All of the Tm values of the FasL binding lowering variants were not significantly different from those of the chimeric A-g4PEK, so it was confirmed that there was no effect on thermostability by introducing 1 or 2 amino acid substitutions into the CRD portion of DcR3 (Fig. 26). ).

[Example 18] Evaluation of in vivo kinetics of DcR3 variants

Chimeric A-Fc (Eg1S) and FasL binding lowering variants E57K-Fc, E57L-Fc, E57R-Fc, E57V-Fc, 45-10-Fc, 45-18-Fc, 82-5-Fc (all Fc for g4PEK), the in vivo kinetics in mice was evaluated in the same manner as in Example 6. Chimeric A-Fc (Eg1S) is produced by stable expression of CHO-K1 cells, each FasL-binding reduced variant is produced by transient expression of CHO-S cells, and SEC (Superdex 200) using AKTApurifier (GE Healthcare) What was purified to 95% or more of monomers by Increase 10/300 GL) (GE Healthcare) was used.

A single dose of 10 mg/kg of each DcR3 variant to 5- to 6-week-old BALB/c mice (♀) i. v. AUC0-∞ (μg*h), which is the area under the concentration-time curve until infinity, and the half-life (h) of the phase of elimination after a single administration (n=2 or 3) when administered (n=2 or 3) /mL) was calculated (Fig. 27). Each DcR3 variant prepared by transient expression of Expi293 cells was used as a standard material for measuring the concentration of the DcR3 variant in the serum of mice administered with E57K-Fc, E57R-Fc, and E57V-Fc.

As a result, compared with S195-Fc (IEGRMD g1S), which is a wild-type DcR3 described in Example 6, the AUC of any DcR3 variant was significantly improved.

[Example 19] In vivo efficacy evaluation of DcR3 variants

In vivo efficacy evaluation of chimeric A-Fc (g4PEK) was performed using a mouse acute xenograft versus host disease (GVHD) model.

(1) Construction of a mouse acute heterogeneous GVHD model

A mouse acute heterogeneous GVHD model was prepared in the same manner as described in JP5209625. Using 6-week-old, severe combined immunodeficiency (SCID) cancer-type mice, 100 μg of rat anti-mouse IL2 receptor-β (IL2Rβ) chain antibody TMβ1 (Bio X Cell) was intraperitoneally injected on days -2 and 5. , depleted endogenous mouse natural killer cells. On the -1 day, a lethal dose of 1.7 Gy or less was irradiated to mice using a CellRad type X-ray irradiation device (Faxitron). On day 0, 3×10 ⁶ human PBMCs (AllCells) were intraperitoneally transferred, and then 300 μg of chimeric A-Fc or DNP antibody (all g4PEK) adjusted with 100 μL of PBS was intraperitoneally injected. For the chimeric A-Fc administration group, 300 μg of chimeric A-Fc was additionally administered on the 4th and 8th days. After 12 days, while determining the gross pathological score by the GVHD reaction, spleens were collected from slaughtered mice, and the number of human cells in the spleens was evaluated.

(2) evaluation of the GVHD condition score

The gross condition observed on day 12 was scored by four indicators: hair shape, redness in the intestinal tract, activity status, and weight loss ( FIG. 28A ). For each index, none, mild, and severe were scored as 0, 1, and 2, respectively, and the scores of all indexes were summed to determine the GVHD condition score.

As a result, an increase in the GVHD condition score due to the translocation of human PBMC and a decrease in the condition score by administration of the chimeric A-Fc compared to the DNP antibody administration group were confirmed, respectively, and the drug efficacy of the chimeric A-Fc was confirmed (Fig. 28b).

(3) Measurement of the number of human cells in the spleen

A mouse spleen was collected, and the spleen was disrupted using gentleMACS Dissociators (Miltenyi) to prepare a spleen cell suspension. After hemolysis with lysing buffer (BD Biosciences), PE/Cy7-labeled anti-human CD45 antibody, FITC-labeled anti-human CD3 antibody, APC-labeled anti-human CD4 antibody, and PE-labeled anti-human CD8 antibody (all manufactured by BioLegend) were stained. , Each human cell subset was analyzed by flow cytometry analysis. CountBright Absolute Counting Beads, for flow cytometry (Thermo Fisher Scientific) were used for the measurement of the number of cells, and the abundance ratio of the count of fluorescent beads detected by flow cytometry and the number of each cell subset was determined by the known amount of added beads. After correction, the total number of cells in the spleen was calculated.

As a result, human cells were detected in the mouse spleen by translocation of human PBMC, and a decrease in the number of human cells in the mouse spleen was confirmed by administration of the chimeric A-Fc compared to the DNP antibody-administered group (FIG. 29).

[Example 20] Evaluation of binding activity to soluble DcR3 ligand trimer

With respect to the chimeric A-Fc and FasL binding lowering variants having various mutated Fc prepared in Example 12, soluble human LIGHT trimer, soluble cynomolgus monkey LIGHT trimer, soluble human TL1A trimer, soluble type Binding activity to cynomolgus monkey TL1A trimer, soluble human FasL trimer, or soluble cynomolgus monkey FasL trimer was evaluated as follows, with some modifications of the methods described in Examples 7 and 13.

(1) Preparation of soluble DcR3 ligand trimer

The human soluble DcR3 ligand trimer prepared in Example 13 was used. Soluble recombinant LIGHT of cynomolgus monkey is a sequence (FLAG-) in which a FLAG tag (DYKDDDDK) is added to the N-terminus, and the extracellular region (Asp74-Val240) (SEQ ID NO: 134) of cynomolgus LIGHT is added downstream thereof (FLAG- cynoLIGHT) (base sequence: SEQ ID NO: 341, amino acid sequence: SEQ ID NO: 342). The same sequence as in Example 7 was used for cynomolgus monkey soluble recombinant TL1A (His6-cynoTL1A) and cynomolgus monkey soluble FasL (His6-cynoFasL) (amino acid sequences SEQ ID NOs: 122 and 124, respectively). LIGHT and TL1A were transiently expressed by Expi293 and FasL by CHO-S.

FLAG-cynoLIGHT and His6-cynoTL1A were purified by the method described in Example 13. For His6-cynoFasL, a washing buffer (60 mM imidazole, 20 mM phosphate) prepared by passing the culture supernatant through a column filled with Ni Sepharose Fast Flow resin (GE Healthcare) and using His Buffer Kit (GE Healthcare) After washing with sodium, 0.5 M NaCl, pH 7.4), it was eluted with an elution buffer (250 mM imidazole, 20 mM sodium phosphate, 0.5 M NaCl, pH 7.4), and the buffer was replaced with PBS.

Purified FLAG-cynoLIGHT and His6-cynoTL1A were fractionated as trimer fractions by gel filtration chromatography (SEC) (Superdex 200 Increase 10/300 GL) (GE Healthcare) using AKTApurifier (GE Healthcare). . Purified His6-cynoFasL was analyzed by SEC-UHPLC (Apparatus: Nexera X2 (Shimazu Seisakusho), Column: ACQUITY UPLC Protein BEH SEC 200 Å, 1.7 µm, 4.6×150 mm (Waters)), 85% or more 3 It was confirmed that it was a monomer.

(2) Measurement of binding activity using BIAcore

As chimeric A-Fc and FasL binding lowering variants having various mutated Fc, all of the variants described in Example 12, SEC (Superdex 200 Increase 10/300 GL) using AKTApurifier (GE Healthcare) (GE Health Care) or SEC (HiLoad 26/600 Superdex 200 pg) (GE Healthcare) using AKTApure 25 (GE Healthcare) purified to 95% or more of monomers was used.

The binding activity to human or cynomolgus monkey DcR3 ligand trimer was analyzed by SPR method. BIAcore T-100 (GE Healthcare) was used to measure binding activity to human LIGHT, cynomolgus monkey LIGHT, human TL1A, and cynomolgus monkey TL1A, and BIAcore T-200 was used to measure binding activity to human FasL and cynomolgus monkey FasL. (GE Healthcare) was used, respectively. HBS-EP+buffer was used as the buffer.

Various DcR3 variants were flowed into Series S Sensor Chip Protein A (GE Healthcare) at 10 μL/min for 30 seconds, and captured. On the other hand, a buffer solution containing no protein was flowed into the reference flow cell. Thereafter, each human DcR3 ligand trimer or each cynomolgus DcR3 ligand trimer diluted to 0.08-20 nmol/L was flowed as an analyte. Human LIGHT trimer, human TL1A trimer, or each cynomolgus monkey DcR3 ligand trimer was flowed for 2 minutes at 30 μL/min to monitor binding, followed by a flow of buffer for 5 minutes to monitor dissociation. Human FasL trimer was flowed at 30 μL/min for 1 minute to monitor binding, followed by a flow of buffer for 3 minutes to monitor dissociation. Next, 3 mol/L magnesium chloride was flowed at 30 µL/min for 1 minute to perform a regeneration reaction. BIAcore T-100 evaluation software for measuring binding activity to human LIGHT, cynomolgus LIGHT, human TL1A, cynomolgus TL1A, 1:1 binding model, BIAcore T- for measuring binding activity to human FasL, cynomolgus FasL Using 200 evaluation software, a 1:1 binding model, each DcR3 ligand as a trimer (human LIGHT trimer: 62.4 kDa, cynomolgus monkey LIGHT trimer: 57.9 kDa, human TL1A trimer: 66.2 kDa, cynomolgus monkey) TL1A trimer: 66.1 kDa, human FasL trimer: 53.1 kDa, cynomolgus FasL trimer: 53.1 kDa), angular kinetic constants (k _a , k _d , K _D ) were calculated.

As a result, the chimeric A-Fc (Eg1S, Eg1S YTE, Eg1S N434A, Eg1S LALAGA, Eg1S LALAGANA) having various mutated Fc (each amino acid sequence: SEQ ID NOs: 168, 170, 172, 176, 178) were both human and human. It was confirmed that binding to each DcR3 trimer ligand of the omnivore monkey (FIGS. 30a, 30b).

In FasL binding lowering variants E57K-Fc, E57R-Fc, E57V-Fc, 45-10-Fc, 45-18-Fc (all Fc is Eg1S YTE, Eg1S N434A or Eg1S LALAGANA) having various mutated Fc, all There was no significant difference in K _D values for human LIGHT trimer and human TL1A trimer compared to chimeric A-Eg1S. However, in all of these FasL binding lowering variants, the K _D value for the human FasL trimer was significantly increased compared to that of the chimeric A-Eg1S, and it was confirmed that the human FasL binding selectively decreased (FIG. 30a) .

In all of the FasL binding lowering variants having various mutated Fc, the K _D value for the cynomolgus monkey LIGHT trimer or the cynomolgus monkey TL1A trimer was about the same as that of the chimeric A-Eg1S, but E57R-Fc (Eg1S YTE, Eg1S N434A) showed a tendency to decrease the binding activity to cynomolgus monkey TL1A trimer. On the other hand, in any of the FasL binding lowering variants, the K _D value for the cynomolgus monkey FasL trimer significantly increased compared to the chimeric A-Eg1S, confirming that FasL-selective binding was reduced (Fig. 30b). ).

[Example 21] Evaluation of neutralizing activity of DcR3 soluble ligand

Neutralizing activity evaluation for soluble human LIGHT, soluble human TL1A, and soluble human FasL of FasL binding lowering variants having various mutated Fc was carried out in the same manner as in Example 9 or Example 14. As various DcR3 variants, chimeric A-Fc prepared in Example 12, or FasL binding lowering variant E57K-Fc, 45-18-Fc produced by transient expression of CHO-S cells (all Fc are Eg1S YTE) or Eg1S LALAGANA), purified to 95% or more of monomers by the method described in (1) of Example 14 or SEC (HiLoad 16/600 Superdex 200 pg) (GE Healthcare) using AKTApurifier (GE Healthcare) one was used.

(1) Soluble human LIGHT neutralizing activity

By the method described in Example 14, the inhibitory activity of various DcR3 variants on LIGHT-dependent CXCL10 production by IFN-γ-stimulated long myofibroblasts was evaluated. As a result, it was confirmed that the chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) inhibited CXCL10 production in a concentration-dependent manner and had neutralizing activity for soluble LIGHT. became The results are shown in FIG. 31 .

(2) Soluble human TL1A neutralizing activity

By the method described in Example 9, the inhibitory activity of various DcR3 variants on TL1A-dependent IFN-γ production by IL-12 and IL-18-stimulated human T cells was evaluated. As a result, the chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) inhibit IFN-γ production in a concentration-dependent manner, and have neutralizing activity against soluble TL1A it was confirmed The results are shown in FIG. 32 .

(3) Soluble human FasL neutralizing activity

By the method described in Example 9, the inhibitory activity of various DcR3 variants against FasL-dependent cell death of Jurkat cells was evaluated. As a result, it was confirmed that the chimeric A-Fc (Eg1S YTE, Eg1S LALAGANA) inhibited the cell death of Jurkat cells in a concentration-dependent manner, and had a neutralizing activity against soluble FasL. On the other hand, E57K-Fc and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) have significantly reduced inhibitory activity against Jurkat cell death, and neutralizing activity for soluble FasL is selectively reduced. it was confirmed The results are shown in FIG. 33 .

In addition, in FIGS. 31 to 33 , the solid line and X (-X-) are the negative control anti-DNP antibody, the solid line and the black circle (-●-) are the chimeric A-Fc (Eg1S YTE), the solid line and the white triangle ( -Δ-) indicates E57K-Fc (Eg1S YTE), solid and white circles (-○-) indicate 45-18-Fc (Eg1S YTE), and dotted and black circles (--●--) indicate chimeric A-Fc ( Eg1S LALAGANA), dashed lines and white triangles (--Δ--) indicate E57K-Fc (Eg1S LALAGANA), and dotted lines and white circles (--○--) indicate 45-18-Fc (Eg1S LALAGANA).

[Example 22] Evaluation of binding activity to DcR3 membrane ligand

For the chimeric A and FasL binding lowering variants having various mutated Fc, the binding activity evaluation for membranous human LIGHT, membranous human TL1A and membranous human FasL was carried out in the same manner as in Example 15.

Chimeric A-Fc (Eg1S) was prepared from a stable expression strain of CHO cells and purified to 95% or more of monomers by SEC was used. Chimeric A-Fc (Eg1S YTE, Eg1S N434A, Eg1S LALAGA, Eg1S LALAGANA) were all prepared by transient expression of CHO-S cells and purified by SEC to 95% or more of the monomers were used. FasL binding lowering variants E57K-Fc, E57R-Fc, E57V-Fc, 45-10-Fc and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) are all produced by transient expression of Expi293F cells , those purified to 95% or more of monomers by SEC were used. SEC was performed in the same manner as in Example 21.

A cell line forcibly expressing a membrane molecule of human LIGHT, human TL1A, or human FasL, respectively, described in Example 15 (also referred to as LIGHT/HEK293, TL1A/CHO, or FasL/CHO, respectively), and its host cell HEK293; The sample was reacted with CHO-K1 cells at a final concentration of 10 μg/mL, secondary antibody Goat F(ab’) ₂ Anti-Human IgG R-phycoerythrin Conjugate (Southern Biotech) at a final concentration of 100 ng/mL, Fluorescence intensity of PE was analyzed by flow cytometer. The results are shown in Figs. 34A, 34B and 34C. The black bar in FIG. 34A shows the reactivity to LIGHT/HEK293, and the white bar shows the reactivity to HEK293 cells. The black bar in FIG. 34B represents the reactivity to TL1A/CHO, and the white bar represents the reactivity to CHO-K1 cells. The black bar in FIG. 34c represents the reactivity to FasL/CHO, and the white bar represents the reactivity to CHO-K1 cells.

As a result, as shown in FIGS. 34A and 34B, for membranous human LIGHT and membranous TL1A, chimeric A-Fc (Eg1S, Eg1S YTE, Eg1S N434A, Eg1S LALAGA, Eg1S LALAGANA), E57K-Fc, E57R-Fc, E57V-Fc, 45-10-Fc and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) were confirmed to bind. On the other hand, as shown in Fig. 34c, with respect to membrane FasL, it was confirmed that the binding activity of any of the FasL binding lowering variants was significantly reduced compared to the chimera A having various mutated Fc.

[Example 23] Evaluation of neutralizing activity of DcR3 membrane ligand

By the method shown below, the neutralizing activity of chimeric A and FasL binding lowering variants having various mutated Fc against membranous human LIGHT, membranous human TL1A and membranous human FasL was evaluated. Chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) were all prepared by transient expression of CHO-S, and 95% or more of monomers were used by SEC in the same manner as in Example 21. The purified one was used.

(1) Membrane-type human LIGHT neutralizing activity

CXCL10 production by co-culture with the membranous LIGHT forced expression HEK293 cell line (LIGHT/HEK293) used in Example 22 using human long myofibroblasts (Lonza) as an index, chimeric A-Fc, E57K-Fc, 45- The membrane-type human LIGHT neutralizing activity of 18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) was evaluated.

Human intestinal myofibroblasts were seeded at 1×10 ⁴ cells/well in a Collagen I coat 96 well plate (Corning), and cultured at 37° C. overnight. After fixing with Fixation Buffer (BD Bioscience), LIGHT/HEK293 washed with PBS and medium were 1×10 ⁴ cells/well, and recombinant human IFN-γ (Fujifilm Wako Junyaku) was used at a final concentration of 10ng/mL , chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) were added to a final concentration of 4.88, 19.5, 78.1, 313, 1250 or 5000 ng/mL, respectively, and the total amount of the culture solution It prepared so that it might become this 200 microliters/well. The culture supernatant after culturing for 4 days was collected, and the CXCL10 concentration in the culture supernatant was measured using CXCL10/IP-10 (human) AlphaLISA Detection Kit (Perkin Elmer). The results are shown in FIG. 35 . In Figure 35, the solid line and X (-X-) are the negative control anti-DNP antibody, the solid line and the black circle (-●-) are the chimeric A-Fc (Eg1S YTE), the solid line and the white triangle (-Δ-) are E57K-Fc (Eg1S YTE), solid and white circles (-○-) are 45-18-Fc (Eg1S YTE), dotted and black circles (--●--) are chimeric A-Fc (Eg1S LALAGANA), dotted lines and white triangles (--Δ--) indicate E57K-Fc (Eg1S LALAGANA), and dotted lines and white circles (--○--) indicate 45-18-Fc (Eg1S LALAGANA).

As shown in Fig. 35, CXCL10 production by membrane-type LIGHT was inhibited in a concentration-dependent manner with respect to any of chimeras A-Fc, E57K-Fc, and 45-18-Fc (Fc is Eg1S YTE or Eg1S LALAGANA), and the chimera A and FasL binding lowering variants E57K-Fc and 45-18-Fc were both confirmed to have a membrane-type LIGHT neutralizing activity.

(2) Membrane-type human TL1A neutralizing activity

Chimeric A-Fc, E57K-Fc, 45 using human CD4-positive T cells and co-culturing with the membranous TL1A forced expression CHO-K1 cell line (TL1A/CHO) used in Example 22 as an index for IFN-γ production Membrane-type human TL1A neutralizing activity of -18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) was evaluated.

CD4 positive T cells were isolated from frozen healthy PBMCs (AllCells) using EasySep Human CD4+ T cell Isolation Kit (STEMCELL Technologies). The isolated CD4-positive T cells were seeded at 2×10 ⁵ cells/well in a 96-well floating culture plate, and then human IL-12 (Miltenyi Biotech) was added to a final concentration of 2 ng/mL and Recombinant Human IL-18 (MBL). ) was added to a final concentration of 50 ng/mL, and Human IL-15 (Miltenyi Biotech) was added to a final concentration of 25 ng/mL, and fixed with Fixation Buffer (BD Biosciences). Thereafter, 3 × 10 ⁴ cells/well of membrane-type TL1A-expressing CHO-K1 cells washed with PBS and medium, chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) Each was added so as to have a final concentration of 20, 50, 100, 250, 500, or 1000 ng/mL, and the total amount of the culture solution was prepared to be 200 µL/well. The culture supernatant after culturing for 2 days was collected, and the IFN-γ concentration in the culture supernatant was measured using AlphaLISA IFN-γ Immunoassay Research Kit (Perkin Elmer). The results are shown in FIG. 36 . In Figure 36, the solid line and X (-X-) are the negative control anti-DNP antibody, the solid line and the black circle (-●-) are the chimeric A-Fc (Eg1S YTE), the solid line and the white triangle (-Δ-) are E57K-Fc (Eg1S YTE), solid and white circles (-○-) are 45-18-Fc (Eg1S YTE), dotted and black circles (--●--) are chimeric A-Fc (Eg1S LALAGANA), dotted lines and white triangles (--Δ--) indicate E57K-Fc (Eg1S LALAGANA), and dotted lines and white circles (--○--) indicate 45-18-Fc (Eg1S LALAGANA).

As shown in Fig. 36, the production of IFN-γ by membrane type TL1A was inhibited in a concentration-dependent manner for any of the chimeric A-Fc, E57K-Fc, and 45-18-Fc (Fc is Eg1S YTE or Eg1S LALAGANA). , it was confirmed that all of the FasL binding-reducing variants E57K-Fc and 45-18-Fc maintained the same membrane-type TL1A neutralizing activity as that of the chimeric A-Fc.

(3) Membrane-type human FasL neutralizing activity

Membrane-type FasL forced-expressing strain in which the neutralizing activity of chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) against membrane-type human FasL as a soluble type is not cleaved Apoptosis of Jurkat cells by co-culture with

A FLAG tag (DYKDDDDK) was added to the N-terminus, and a FasL cleavage site (Glu128-Ile131) (Nat. Med., 1998, 4(1), p.31-36) and a proline-rich region of the intracellular region (Pro8-) Pro69) (Nat. Med., 1996, 2(3), p.317-322) was removed, a plasmid expressing the membranous human FasL variant sequence (base sequence: SEQ ID NO: 343, amino acid sequence: SEQ ID NO: 344) It was produced by the following method. Using the membrane-type FasL expression plasmid described in Example 15 as a template and using primers designed to delete the target sequence, PCR was performed for three regions from PstI site to Tyr7, Pro70 to Leu127, and from Gly132 to NotI site, respectively. The amplified product was inserted under the CMV promoter of the pCIpuro vector in the same manner as in Example 1. The obtained plasmid was introduced into CHO-K1 cells in the same manner as in Example 15, and the membrane-type human FasL mutant high expression CHO-K1 cell line (FasLdel) by cell sorting of the positive fraction of the PE-labeled anti-human FasL antibody (BioLegend). /CHO) was obtained.

FasLdel/CHO was seeded at 5×10 ³ cells/well in a 96-well adhesion culture plate (Sumitomo Bakelite) and cultured overnight at 37° C. in a 5% CO ₂ incubator, and then the cells were washed once. Here, Jurkat cells were seeded at 5×10 ⁴ cells/well, and chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc Eg1S YTE or Eg1S LALAGANA) was added. The total amount of the culture medium was adjusted to 100 µL/well, and after culturing for 4 hours at 37° C. in a 5% CO ₂ incubator, only Jurkat cells were recovered. The recovered Jurkat cells were seeded in 96-well U-bottom plates (Falcon) and stained for 15 minutes at room temperature using a PE annexin V apoptosis detection kit (BD Biosciences). After washing the cells with a buffer for FCM, the fluorescence intensity was analyzed with a flow cytometer FACS Fortessa (BD Biosciences), and Annexin V-positive cells were detected as dead cell fractions. The results are shown in FIG. 37 . In Figure 37, the solid line and X (-X-) are the negative control anti-DNP antibody, the solid line and the black circle (-●-) are the chimeric A-Fc (Eg1S YTE), the solid line and the white triangle (-Δ-) are E57K-Fc (Eg1S YTE), solid and white circles (-○-) are 45-18-Fc (Eg1S YTE), dotted and black circles (--●--) are chimeric A-Fc (Eg1S LALAGANA), dotted lines and white triangles (--Δ--) indicate E57K-Fc (Eg1S LALAGANA), and dotted lines and white circles (--○--) indicate 45-18-Fc (Eg1S LALAGANA).

As shown in FIG. 37 , it was confirmed that the chimeric A-Fc (Eg1S YTE, Eg1S LALAGANA) inhibited the cell death of Jurkat cells in a concentration-dependent manner, and had a neutralizing activity against membrane FasL. On the other hand, with respect to E57K-Fc and 45-18-Fc (Fc is either Eg1S YTE or Eg1S LALAGANA), both showed little inhibitory activity against Jurkat cell death even in the high concentration addition group of 25000 ng/mL, and neutralizing activity against membrane-type FasL It was confirmed that this remarkably decreased.

[Example 24] Evaluation of binding activity to soluble DcR3 ligand derived from human primary cells

The binding activity of the chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) to various soluble DcR ligands was evaluated using soluble DcR3 ligands derived from human primary cells. did For any of the test substances, those prepared by transient expression of CHO-S and purified to 95% or more of monomers by SEC were used in the same manner as in Example 21.

(1) Primary LIGHT binding activity

Binding of the chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) to primary soluble LIGHT produced from activated human T cells was evaluated by the following method. Primary soluble LIGHT was prepared by the following method. 7 x 10 human T cells isolated from thawed frozen healthy PBMCs (AllCells) in a 6-well adherent culture plate (Sumitomo Bakelite) immobilized with 10 µg/mL anti-CD3 antibody OKT3 (BioLegend) immobilized After seeding at ⁶ cells/well, an anti-CD28 antibody (BioLegend, Inc.) with a final concentration of 1 μg/mL was added and cultured in a 5% CO ₂ incubator for 3 days, and then the culture supernatant was recovered. The primary soluble LIGHT concentration in the recovered culture supernatant was measured using Human LIGHT/TNFSF14 Quantikine ELISA Kit (R&D), and 14 pg/mL in the culture supernatant of unstimulated human T cells, anti-CD3 antibody and anti-CD28 antibody It was confirmed that 5.6 ng/mL of soluble LIGHT was present in the culture supernatant of human T cells stimulated with

Next, 10 μg/mL anti-human IgG antibody (American Qualex) was immobilized on a 96-well immunization plate (Thermo Scientific) and blocked with 2% Block Ace (DS Pharma Biomedical), followed by 2 μg /mL of chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA). The culture supernatant of human T cells induced to produce light was concentrated 6-fold using Amicon Ultra-15 (Millipore) with a molecular weight cutoff of 3 kDa pre-rinsed with sterile water, and the plate was washed and then concentrated 6-fold. The supernatant was reacted. As a standard for LIGHT, the soluble recombinant FLAG-LIGHT trimer prepared in Example 13 was used. After washing the plate, a biotinylated anti-LIGHT antibody (R&D) was reacted. After washing the plate, react with streptavidin-HRP (PIERCE), wash again, add TMB solution (Dako) to develop color, stop the color reaction with 2N sulfuric acid solution, 450 nm (reference The absorbance at a wavelength of 570 nm) was measured.

Fig. 38A shows the results of the soluble recombinant FLAG-LIGHT trimer, and Fig. 38B shows the results of the culture supernatant of LIGHT-induced human T cells. In Figure 38a, the solid line and X (-X-) are the negative control anti-DNP antibody, the solid line and the black circle (-●-) are the chimeric A-Fc (Eg1S YTE), the solid line and the white square (-□-) are Chimeric A-Fc (Eg1S), solid and white triangles (-Δ-) are E57K-Fc (Eg1S YTE), solid and white circles (-○-) are 45-18-Fc (Eg1S YTE), dashed and black circles (--●--) is chimeric A-Fc (Eg1S LALAGANA), dashed and white triangles (--△--) are E57K-Fc (Eg1S LALAGANA), dashed and white circles (--○--) are 45 -18-Fc (Eg1S LALAGANA). In Fig. 38B, "6 x sup. "CD3/28" is a culture supernatant of 6-fold concentrated human T cells induced with LIGHT expression under anti-CD3 antibody OKT3 and anti-CD28 antibody stimulation conditions. It shows that the culture supernatant concentrated 6-fold of unstimulated human T cells was used.

As shown in FIG. 38B , human T induced LIGHT expression also in wells in which any of chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S, Eg1S YTE, or Eg1S LALAGANA) were captured. LIGHT was detected only in the culture supernatant of the cells. In this regard, chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S, Eg1S YTE, or Eg1S LALAGANA) are soluble from human T cells stimulated with anti-CD3 antibody and anti-CD28 antibody. It was confirmed to bind to the type LIGHT.

(2) primary TL1A binding activity

Binding of the chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) to activated human PBMC-derived primary soluble TL1A was evaluated. Primary soluble TL1A was prepared by the following method. A human IgG antibody (Jackson ImmunoResearch) at 250 µg/mL was immobilized on a 6-well adhesion culture plate (Sumitomo Bakelite), and an anti-human κ light chain antibody (Bethyl Laboratories) and an anti-human λ light chain antibody (Bethyl Laboratories) were also immobilized. g) were added to a final concentration of 100 μg/mL, and treated for 1 hour to prepare an immune complex. On the plate, thawed frozen healthy PBMCs (AllCells) were seeded at 6×10 ⁶ cells/well, and cultured at 37° C. in a 5% CO ₂ incubator. After 3 days, the culture supernatant was recovered and concentrated 4 times using Amicon Ultra-15 (Millipore) having a molecular weight cutoff of 3 kDa, pre-rinsed with sterile water.

Sandwich ELISA was performed in the same manner as in (1). Chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) or the anti-human TL1A antibody prepared in Example 16 were captured in 96-well immune plates. Next, the plate was reacted with a 4-fold concentrated culture supernatant of PBMC induced to produce TL1A, or the soluble recombinant His6-TL1A trimer prepared in Example 13 as a standard, followed by a biotinylated anti-TL1A antibody (R&D) detected.

Fig. 38C shows the result of the soluble recombinant His6-TL1A trimer, and Fig. 38D shows the result of the PBMC culture supernatant induced to produce TL1A. In Figure 38c, solid line and X (-X-) are negative control anti-DNP antibody, solid line and black circle (-●-) are chimeric A-Fc (Eg1S YTE), and solid white square (-□-) is Chimeric A-Fc (Eg1S), solid and white triangles (-Δ-) are E57K-Fc (Eg1S YTE), solid and white circles (-○-) are 45-18-Fc (Eg1S YTE), dashed and black circles (--●--) is chimeric A-Fc (Eg1S LALAGANA), dashed and white triangles (--△--) are E57K-Fc (Eg1S LALAGANA), dashed and white circles (--○--) are 45 -18-Fc (Eg1S LALAGANA), solid black triangles (-▲-) indicate anti-TL1A antibody. In FIG. 38D , “4×sup.I.C.” indicated by a black bar is a culture supernatant of PBMC induced to induce TL1A expression under immune complex stimulation conditions, and “4×sup.I.C.” indicated by a white bar is “no” , indicates that an unstimulated PBMC culture supernatant was used. As shown in FIG. 38D , even for wells in which any of the chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S, Eg1S YTE, or Eg1S LALAGANA) were captured, PBMCs inducing TL1A expression were TL1A was detected only in the culture supernatant. In this regard, it was confirmed that the chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc are Eg1S, Eg1S YTE, or Eg1S LALAGANA) bind to soluble TL1A produced from PBMCs stimulated with immune complexes. became

(3) primary FasL binding activity

Binding of the primary chimeric A-Fc, E57K-Fc, and 45-18-Fc (all Fc is Eg1S YTE or Eg1S LALAGANA) to soluble human FasL was evaluated. As the primary soluble FasL, soluble FasL produced by human T cells induced by activation-induced cell death (AICD) was prepared in the same manner as in Example 16 and used.

Sandwich ELISA was performed in the same manner as in (1). Chimeric A-Fc, E57K-Fc, 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA), or Fas-Fc (R&D) were captured in a 96-well immune plate, followed by AICD induced human T Cells were reacted with a 6-fold concentrated culture supernatant, or Human His6 Fas Ligand/TNFSF6 (Cell Signaling TECHNOLOGY) as a standard, and detected with a biotinylated anti-FasL antibody supplied with the Human Fas Ligand ELISA Kit (Abcam).

Fig. 38E shows the results of soluble recombinant His6-FasL, and Fig. 38F shows the results of the culture supernatant of AICD-induced human T cells. In Figure 38e, solid line and X (-X-) are negative control anti-DNP antibody, solid line and black circle (-●-) are chimeric A-Fc (Eg1S YTE), solid line and white square (-□-) are Chimera A-Fc (Eg1S), solid line and white triangle (-Δ-) is E57K-Fc (Eg1S YTE), solid line and white circle 「-○-」 is 45-18-Fc (Eg1S YTE), dotted line and black circle (--●--) is chimeric A-Fc (Eg1S LALAGANA), dashed and white triangles (--△--) are E57K-Fc (Eg1S LALAGANA), dashed and white circles (--○--) are 45 -18-Fc (Eg1S LALAGANA), solid lines and black triangles (-▲-) represent Fas-Fc. In Fig. 38F, "6xsup.CD3" indicated by black bars is a culture supernatant of human T cells induced with AICD under anti-CD3 antibody OKT3 stimulation conditions, and "6xsup.CD3" indicated by white bars. "None" indicates that the culture supernatant of human T cells not induced with AICD was used, respectively. As shown in Fig. 38F, in the wells in which the chimeric A-Fc (Eg1S, Eg1S YTE, Eg1S LALAGANA) were captured, FasL was detected only in the culture supernatant of AICD-induced human T cells. On the other hand, in wells in which E57K-Fc and 45-18-Fc (both Fc are Eg1S YTE or Eg1S LALAGANA) were captured, FasL detected in the culture supernatant of AICD-induced human T cells was significantly reduced. It was confirmed that the mutant had reduced binding activity to human T-cell-derived soluble FasL.

[Example 25] Evaluation of physical properties of DcR3 variants

E57K-Fc and 45-18-Fc (all Fc are Eg1S YTE or Eg1S LALAGANA) prepared by transient expression of CHO-S and purified to 95% or more of monomers by SEC in the same manner as in Example 21, Examples In the same manner as in 4, the elution time (min) was calculated by hydrophobic interaction chromatography (HIC) and the Tm value (°C) was calculated by the differential scanning fluorescence (DSF) method.

As a result, since none of the values showed values equivalent to those of Example 17, when the introduction of the amino acid mutation into the IgG1 Fc sequence and the amino acid substitution into the chimeric A sequence were combined, hydrophobicity and thermostability were not affected. it was confirmed

[Example 26] Evaluation of in vivo kinetics of DcR3 variants

Chimeric A-Fc, E57K-Fc, 45-18-Fc produced by transient expression of CHO-S and purified to 95% or more of monomers by SEC in the same manner as in Example 21 (all Fc are Eg1S YTE or Eg1S LALAGANA) , the in vivo kinetics in mice was evaluated in the same manner as in Example 6. The results are shown in FIG. 39 .

As shown in FIG. 39 , compared with the AUC value of 354 of S195-Fc (IEGRMD g1S), which is a wild-type DcR3 described in Example 6, the AUC of any DcR3 variant was significantly improved.

[Example 27] Evaluation of aggregation of DcR3 variants not fused to Fc

The C-terminal His6 tag of a human wild-type DcR3 (S195) lacking the HBD region (S195-His6) (base sequence: SEQ ID NO: 345, amino acid sequence: SEQ ID NO: 346), a His6 tag at the C-terminus of chimera A Protein to which was added (chimeric A-His6) (base sequence: SEQ ID NO: 347, amino acid sequence: SEQ ID NO: 348), a protein with a His6 tag added to the C terminus of chimeric A-E57K (chimeric A-E57K-His6) (base Sequence: SEQ ID NO: 349, amino acid sequence: SEQ ID NO: 350), a protein with a His6 tag added to the C terminus of 45-18 (45-18-His6) (base sequence: SEQ ID NO: 351, amino acid sequence: SEQ ID NO: 352) It produced as follows.

S195-His6 was artificially synthesized (Pasmax) by adding a signal peptide sequence DNA fragment to the N-terminus and a His6 tag sequence to the C-terminus of a human DcR3 DNA fragment (SEQ ID NO: 107) excluding HBD. Chimera A-His6, chimeric A-E57K-His6, 45-18-His6 are, respectively, the DNA fragment of chimera A (SEQ ID NO: 353), the DNA fragment of chimera A-E57K (SEQ ID NO: 354), or the DNA of 45-18 A DNA fragment of the signal peptide sequence at the N-terminus of the fragment (SEQ ID NO: 355) and a His6 tag sequence at the C-terminus were artificially synthesized. Each artificially synthesized fragment was introduced into the pCIpuro vector by the method described in Example 1, and transient expression was performed by Expi293 cells. Using Amicon Ultra-0.5 (Millipore) with a molecular weight cutoff of 3 kDa pre-rinsed with sterile water, the culture supernatant expressing S195-His6 was concentrated 30-fold, chimera A-His6, chimera A-E57K-His6, 45 Each of the culture supernatants expressing -18-His6 was diluted 6-fold.

Using these culture supernatants, SDS-PAGE was performed under non-reducing conditions or under reducing conditions of 100 mM DTT, and then a mouse anti- 6x-His-tag monoclonal antibody (Thermo Scientific) was added to the primary antibody, and the secondary antibody was used. Immunoblot was performed using a goat anti-mouse IgG antibody (Southern Biotech) as the antibody.

The results are shown in FIG. 40 . In the culture supernatant in which S195-His6 was transiently expressed, a band was detected near the expected molecular weight of 20.9 kDa under the reducing condition of lane 1, but no band of the monomer was detected under the non-reducing condition of lane 5, and the whole was smeared. had been From this, it was confirmed that all of S195-His6 existed as aggregates in the culture supernatant. On the other hand, the culture supernatant transiently expressing chimera A-His6, chimera A-E57K-His6 and 45-18-His6 were all under the non-reducing conditions in lanes 2 to 4, and under the reducing conditions in lanes 6 to 8, similarly to the expected molecular weight 22.2 A band of the monomer was detected near kDa. From this point, it was shown that the DcR3 variant of the present invention has a significantly reduced aggregate content compared with wild-type DcR3. That is, by substituting CRD1 and CRD4 of DcR3 with CRD1 and CRD4 derived from OPG, respectively, there is an effect of reducing the aggregation seen in wild-type DcR3, and this effect is exhibited regardless of whether the binding to FasL is reduced. appear.

<sequence list free text>

SEQ ID NO: 1: nucleotide sequence of human DcR3cDNA (accession number: NM_003823.3)

SEQ ID NO: 2: Amino acid sequence of human DcR3 (Accession Number: NP_003814.1)

SEQ ID NO: 3: Base sequence of human DcR3 (no signal peptide)

SEQ ID NO: 4: Amino acid sequence of human DcR3 (no signal peptide)

SEQ ID NO: 5: Base sequence of CRD1 of human DcR3

SEQ ID NO: 6: Amino acid sequence of CRD1 of human DcR3

SEQ ID NO: 7: Base sequence of CRD2 of human DcR3

SEQ ID NO: 8: amino acid sequence of CRD2 of human DcR3

SEQ ID NO: 9: Base sequence of CRD3 of human DcR3

SEQ ID NO: 10: amino acid sequence of CRD3 of human DcR3

SEQ ID NO: 11: Base sequence of CRD4 of human DcR3

SEQ ID NO: 12: amino acid sequence of CRD4 of human DcR3

SEQ ID NO: 13: nucleotide sequence of human OPG cDNA (accession number: NM_002546.3)

SEQ ID NO: 14: amino acid sequence of human OPG (accession number: NP_002537.3)

SEQ ID NO: 15: base sequence of human OPG (no signal peptide)

SEQ ID NO: 16: Amino acid sequence of human OPG (no signal peptide)

SEQ ID NO: 17: base sequence of CRD1 of human OPG

SEQ ID NO: 18: amino acid sequence of CRD1 of human OPG

SEQ ID NO: 19: base sequence of CRD2 of human OPG

SEQ ID NO: 20: amino acid sequence of CRD2 of human OPG

SEQ ID NO: 21: base sequence of CRD3 of human OPG

SEQ ID NO: 22: amino acid sequence of CRD3 of human OPG

SEQ ID NO: 23: base sequence of CRD4 of human OPG

SEQ ID NO: 24: Amino acid sequence of CRD4 of human OPG

SEQ ID NO: 25: base sequence of chimeric B-HBD

SEQ ID NO: 26: Amino acid sequence of chimeric B-HBD

SEQ ID NO: 27: base sequence of chimeric C-HBD

SEQ ID NO: 28: Amino acid sequence of chimeric C-HBD

SEQ ID NO: 29: base sequence of chimeric A-HBD

SEQ ID NO: 30: Amino acid sequence of chimeric A-HBD

SEQ ID NO: 31: base sequence of 103-123OPG-HBD

SEQ ID NO: 32: amino acid sequence of 103-123OPG-HBD

SEQ ID NO: 33: base sequence of N131S/N144S-HBD

SEQ ID NO: 34: amino acid sequence of N131S/N144S-HBD

SEQ ID NO: 35: base sequence of T133A/S146A-HBD

SEQ ID NO: 36: amino acid sequence of T133A/S146A-HBD

SEQ ID NO: 37: base sequence of N131S/N144S/N157S-HBD

SEQ ID NO: 38: amino acid sequence of N131S/N144S/N157S-HBD

SEQ ID NO: 39: base sequence of T133A/S146A/T159A-HBD

SEQ ID NO: 40: amino acid sequence of T133A/S146A/T159A-HBD

SEQ ID NO: 41: base sequence of chimeric A-E57K-HBD

SEQ ID NO: 42: Amino acid sequence of chimeric A-E57K-HBD

SEQ ID NO: 43: base sequence of chimeric A-E57L-HBD

SEQ ID NO: 44: Amino acid sequence of chimeric A-E57L-HBD

SEQ ID NO: 45: base sequence of chimeric A-R60K-HBD

SEQ ID NO: 46: Amino acid sequence of chimeric A-R60K-HBD

SEQ ID NO: 47: Base sequence of HBD of wild-type DcR3

SEQ ID NO: 48: Amino acid sequence of HBD of wild-type DcR3

SEQ ID NO: 49: base sequence of chimera B

SEQ ID NO: 50: amino acid sequence of chimera B

SEQ ID NO: 51: base sequence of chimeric C

SEQ ID NO: 52: amino acid sequence of chimeric C

SEQ ID NO: 53: base sequence of chimera A

SEQ ID NO: 54: amino acid sequence of chimera A

SEQ ID NO: 55: base sequence of 103-123OPG

SEQ ID NO: 56: amino acid sequence of 103-123OPG

SEQ ID NO: 57: base sequence of N131S / N144S

SEQ ID NO: 58: amino acid sequence of N131S/N144S

SEQ ID NO: 59: T133A / S146A nucleotide sequence

SEQ ID NO: 60: amino acid sequence of T133A/S146A

SEQ ID NO: 61: base sequence of N131S/N144S/N157S

SEQ ID NO: 62: amino acid sequence of N131S/N144S/N157S

SEQ ID NO: 63: base sequence of T133A/S146A/T159A

SEQ ID NO: 64: amino acid sequence of T133A/S146A/T159A

SEQ ID NO: 65: base sequence of chimeric A-E57K

SEQ ID NO: 66: Amino acid sequence of chimeric A-E57K

SEQ ID NO: 67: base sequence of chimeric A-E57L

SEQ ID NO: 68: Amino acid sequence of chimeric A-E57L

SEQ ID NO: 69: base sequence of chimeric A-R60K

SEQ ID NO: 70: amino acid sequence of chimeric A-R60K

SEQ ID NO: 71: base sequence of Fc (g1S)

SEQ ID NO: 72: amino acid sequence of Fc (g1S)

SEQ ID NO: 73: base sequence of Fc (g4PEK)

SEQ ID NO: 74: amino acid sequence of Fc (g4PEK)

SEQ ID NO: 75: base sequence of chimeric B-Fc (IEGRMD g1S)

SEQ ID NO: 76: Amino acid sequence of chimeric B-Fc (IEGRMD g1S)

SEQ ID NO: 77: base sequence of chimeric C-Fc (IEGRMD g1S)

SEQ ID NO: 78: Amino acid sequence of chimeric C-Fc (IEGRMD g1S)

SEQ ID NO: 79: base sequence of chimeric A-Fc (IEGRMD g1S)

SEQ ID NO: 80: Amino acid sequence of chimeric A-Fc (IEGRMD g1S)

SEQ ID NO: 81: base sequence of chimeric A-Fc (g4PEK)

SEQ ID NO: 82: Amino acid sequence of chimeric A-Fc (g4PEK)

SEQ ID NO: 83: base sequence of 103-123OPG-Fc (g4PEK)

SEQ ID NO: 84: amino acid sequence of 103-123OPG-Fc (g4PEK)

SEQ ID NO: 85: base sequence of N131S/N144S-Fc (g4PEK)

SEQ ID NO: 86: amino acid sequence of N131S/N144S-Fc (g4PEK)

SEQ ID NO: 87: T133A / S146A-Fc (g4PEK) nucleotide sequence

SEQ ID NO: 88: amino acid sequence of T133A/S146A-Fc (g4PEK)

SEQ ID NO: 89: base sequence of N131S/N144S/N157S-Fc (g4PEK)

SEQ ID NO: 90: amino acid sequence of N131S/N144S/N157S-Fc (g4PEK)

SEQ ID NO: 91: base sequence of T133A/S146A/T159A-Fc (g4PEK)

SEQ ID NO: 92: amino acid sequence of T133A/S146A/T159A-Fc(g4PEK)

SEQ ID NO: 93: base sequence of chimeric A-E57K-Fc (g4PEK)

SEQ ID NO: 94: Amino acid sequence of chimeric A-E57K-Fc (g4PEK)

SEQ ID NO: 95: base sequence of chimeric A-E57L-Fc (g4PEK)

SEQ ID NO: 96: Amino acid sequence of chimeric A-E57L-Fc (g4PEK)

SEQ ID NO: 97: base sequence of chimeric A-R60K-Fc (g4PEK)

SEQ ID NO: 98: Amino acid sequence of chimeric A-R60K-Fc (g4PEK)

SEQ ID NO: 99: base sequence of DcR3 FL-Fc

SEQ ID NO: 100: amino acid sequence of DcR3 FL-Fc

SEQ ID NO: 101: base sequence of S195-Fc

SEQ ID NO: 102: amino acid sequence of S195-Fc

SEQ ID NO: 103: base sequence of DcR3 FL-FLAG

SEQ ID NO: 104: amino acid sequence of DcR3 FL-FLAG

SEQ ID NO: 105: nucleotide sequence of linker IEGRMD

SEQ ID NO: 106: amino acid sequence of linker IEGRMD

SEQ ID NO: 107: base sequence of DcR3 (no signal peptide) in which HBD is deleted

SEQ ID NO: 108: amino acid sequence of DcR3 (no signal peptide) with HBD deleted

SEQ ID NO: 109: base sequence of FLAG tag

SEQ ID NO: 110: amino acid sequence of FLAG tag

SEQ ID NO: 111: base sequence of R218Q DcR3

SEQ ID NO: 112: amino acid sequence of R218Q DcR3

SEQ ID NO: 113: nucleotide sequence of human LIGHT soluble recombinant

SEQ ID NO: 114: Amino acid sequence of soluble recombinant of human LIGHT

SEQ ID NO: 115: nucleotide sequence of human TL1A soluble recombinant

SEQ ID NO: 116: Amino acid sequence of soluble recombinant of human TL1A

SEQ ID NO: 117: nucleotide sequence of human FasL soluble recombinant

SEQ ID NO: 118: Amino acid sequence of soluble recombinant of human FasL

SEQ ID NO: 119: base sequence of soluble recombinant of cynomolgus monkey LIGHT

SEQ ID NO: 120: Amino acid sequence of soluble recombinant of cynomolgus monkey LIGHT

SEQ ID NO: 121: base sequence of soluble recombinant of cynomolgus monkey TL1A

SEQ ID NO: 122: Amino acid sequence of soluble recombinant of cynomolgus monkey TL1A

SEQ ID NO: 123: base sequence of soluble recombinant of cynomolgus monkey FasL

SEQ ID NO: 124: Amino acid sequence of soluble recombinant of cynomolgus monkey FasL

SEQ ID NO: 125: Base sequence of soluble recombinant of mouse LIGHT

SEQ ID NO: 126: Amino acid sequence of a soluble recombinant of mouse LIGHT

SEQ ID NO: 127: Base sequence of soluble recombinant of mouse TL1A

SEQ ID NO: 128: amino acid sequence of soluble recombinant of mouse TL1A

SEQ ID NO: 129: base sequence of soluble recombinant of mouse FasL

SEQ ID NO: 130: Amino acid sequence of soluble recombinant of mouse FasL

SEQ ID NO: 131: base sequence of the extracellular region of human LIGHT (Asp74-Val240)

SEQ ID NO: 132: amino acid sequence of the extracellular region of human LIGHT (Asp74-Val240)

SEQ ID NO: 133: base sequence of the extracellular region (Asp74-Val240) of cynomolgus monkey LIGHT

SEQ ID NO: 134: Amino acid sequence of the extracellular region of cynomolgus LIGHT (Asp74-Val240)

SEQ ID NO: 135: base sequence of the extracellular region of mouse LIGHT (Asp72-Val239)

SEQ ID NO: 136: amino acid sequence of the extracellular region of mouse LIGHT (Asp72-Val239)

SEQ ID NO: 137: nucleotide sequence of the extracellular region of human TL1A (Leu72-Leu251)

SEQ ID NO: 138: amino acid sequence of the extracellular region of human TL1A (Leu72-Leu251)

SEQ ID NO: 139: Base sequence of extracellular region (Leu72-Leu251) of cynomolgus monkey TL1A

SEQ ID NO: 140: amino acid sequence of the extracellular region of cynomolgus TL1A (Leu72-Leu251)

SEQ ID NO: 141: base sequence of the extracellular region of mouse TL1A (Ile94-Leu270)

SEQ ID NO: 142: amino acid sequence of the extracellular region of mouse TL1A (Ile94-Leu270)

SEQ ID NO: 143: Base sequence of the extracellular region of human FasL (Pro134-Leu281)

SEQ ID NO: 144: amino acid sequence of the extracellular region of human FasL (Pro134-Leu281)

SEQ ID NO: 145: Base sequence of the extracellular region (Pro133-Leu280) of cynomolgus FasL

SEQ ID NO: 146: Amino acid sequence of the extracellular region of cynomolgus FasL (Pro133-Leu280)

SEQ ID NO: 147: Base sequence of the extracellular region of mouse FasL (Pro132-Leu279)

SEQ ID NO: 148: Amino acid sequence of the extracellular region of mouse FasL (Pro132-Leu279)

SEQ ID NO: 149: base sequence of chimeric A-Fc (g1S)

SEQ ID NO: 150: amino acid sequence of chimeric A-Fc (g1S)

SEQ ID NO: 151: base sequence of OPG K194-Fc

SEQ ID NO: 152: amino acid sequence of OPG K194-Fc

SEQ ID NO: 153: amino acid sequence of constant region of human IgG1 heavy chain

SEQ ID NO: 154: amino acid sequence of constant region of human IgG4 heavy chain

SEQ ID NO: 155: base sequence of Fc (Eg1S)

SEQ ID NO: 156: amino acid sequence of Fc (Eg1S)

SEQ ID NO: 157: base sequence of Fc (Eg1S-YTE)

SEQ ID NO: 158: amino acid sequence of Fc (Eg1S-YTE)

SEQ ID NO: 159: base sequence of Fc (Eg1S-N434A)

SEQ ID NO: 160: amino acid sequence of Fc (Eg1S-N434A)

SEQ ID NO: 161: base sequence of Fc (g1S-LALAGA)

SEQ ID NO: 162: amino acid sequence of Fc (g1S-LALAGA)

SEQ ID NO: 163: base sequence of Fc (Eg1S-LALAGA)

SEQ ID NO: 164: amino acid sequence of Fc (Eg1S-LALAGA)

SEQ ID NO: 165: base sequence of Fc (Eg1S-LALAGANA)

SEQ ID NO: 166: amino acid sequence of Fc (Eg1S-LALAGANA)

SEQ ID NO: 167: base sequence of chimeric A-Fc (Eg1S)

SEQ ID NO: 168: Amino acid sequence of chimeric A-Fc (Eg1S)

SEQ ID NO: 169: base sequence of chimeric A-Fc (Eg1S-YTE)

SEQ ID NO: 170: Amino acid sequence of chimeric A-Fc (Eg1S-YTE)

SEQ ID NO: 171: base sequence of chimeric A-Fc (Eg1S-N434A)

SEQ ID NO: 172: Amino acid sequence of chimeric A-Fc (Eg1S-N434A)

SEQ ID NO: 173: base sequence of chimeric A-Fc (g1S-LALAGA)

SEQ ID NO: 174: Amino acid sequence of chimeric A-Fc (g1S-LALAGA)

SEQ ID NO: 175: base sequence of chimeric A-Fc (Eg1S-LALAGA)

SEQ ID NO: 176: Amino acid sequence of chimeric A-Fc (Eg1S-LALAGA)

SEQ ID NO: 177: base sequence of chimeric A-Fc (Eg1S-LALAGANA)

SEQ ID NO: 178: Amino acid sequence of chimeric A-Fc (Eg1S-LALAGANA)

SEQ ID NO: 179: base sequence of chimeric A-E57R

SEQ ID NO: 180: amino acid sequence of chimeric A-E57R

SEQ ID NO: 181: base sequence of chimeric A-E57V

SEQ ID NO: 182: Amino acid sequence of chimeric A-E57V

SEQ ID NO: 183: base sequence of chimeric A-E57K_R58D

SEQ ID NO: 184: Amino acid sequence of chimeric A-E57K_R58D

SEQ ID NO: 185: base sequence of chimeric A-E57K_R58E

SEQ ID NO: 186: Amino acid sequence of chimeric A-E57K_R58E

SEQ ID NO: 187: base sequence of chimeric A-E57R_R58D

SEQ ID NO: 188: Amino acid sequence of chimeric A-E57R_R58D

SEQ ID NO: 189: base sequence of chimeric A-E57R-Fc (g4PEK)

SEQ ID NO: 190: Amino acid sequence of chimeric A-E57R-Fc (g4PEK)

SEQ ID NO: 191: base sequence of chimeric A-E57V-Fc (g4PEK)

SEQ ID NO: 192: Amino acid sequence of chimeric A-E57V-Fc (g4PEK)

SEQ ID NO: 193: base sequence of chimeric A-E57K_R58D-Fc (g4PEK)

SEQ ID NO: 194: Amino acid sequence of chimeric A-E57K_R58D-Fc (g4PEK)

SEQ ID NO: 195: base sequence of chimeric A-E57K_R58E-Fc (g4PEK)

SEQ ID NO: 196: Amino acid sequence of chimeric A-E57K_R58E-Fc (g4PEK)

SEQ ID NO: 197: base sequence of chimeric A-E57R_R58D-Fc (g4PEK)

SEQ ID NO: 198: Amino acid sequence of chimeric A-E57R_R58D-Fc (g4PEK)

SEQ ID NO: 199: base sequence of chimeric A-E57K-Fc (Eg1S)

SEQ ID NO: 200: Amino acid sequence of chimeric A-E57K-Fc (Eg1S)

SEQ ID NO: 201: base sequence of chimeric A-E57L-Fc (Eg1S)

SEQ ID NO: 202: Amino acid sequence of chimeric A-E57L-Fc (Eg1S)

SEQ ID NO: 203: base sequence of chimeric A-E57R-Fc (Eg1S)

SEQ ID NO: 204: Amino acid sequence of chimeric A-E57R-Fc (Eg1S)

SEQ ID NO: 205: base sequence of chimeric A-E57V-Fc (Eg1S)

SEQ ID NO: 206: Amino acid sequence of chimeric A-E57V-Fc (Eg1S)

SEQ ID NO: 207: base sequence of chimeric A-E57K_R58D-Fc (Eg1S)

SEQ ID NO: 208: Amino acid sequence of chimeric A-E57K_R58D-Fc (Eg1S)

SEQ ID NO: 209: base sequence of chimeric A-E57K_R58E-Fc (Eg1S)

SEQ ID NO: 210: Amino acid sequence of chimeric A-E57K_R58E-Fc (Eg1S)

SEQ ID NO: 211: base sequence of chimeric A-E57R_R58D-Fc (Eg1S)

SEQ ID NO: 212: Amino acid sequence of chimeric A-E57R_R58D-Fc (Eg1S)

SEQ ID NO: 213: base sequence of chimeric A-E57K-Fc (Eg1S YTE)

SEQ ID NO: 214: Amino acid sequence of chimeric A-E57K-Fc (Eg1S YTE)

SEQ ID NO: 215: base sequence of chimeric A-E57L-Fc (Eg1S YTE)

SEQ ID NO: 216: Amino acid sequence of chimeric A-E57L-Fc (Eg1S YTE)

SEQ ID NO: 217: base sequence of chimeric A-E57R-Fc (Eg1S YTE)

SEQ ID NO: 218: Amino acid sequence of chimeric A-E57R-Fc (Eg1S YTE)

SEQ ID NO: 219: base sequence of chimeric A-E57V-Fc (Eg1S YTE)

SEQ ID NO: 220: Amino acid sequence of chimeric A-E57V-Fc (Eg1S YTE)

SEQ ID NO: 221: base sequence of chimeric A-E57K_R58D-Fc (Eg1S YTE)

SEQ ID NO: 222: Amino acid sequence of chimeric A-E57K_R58D-Fc (Eg1S YTE)

SEQ ID NO: 223: base sequence of chimeric A-E57K_R58E-Fc (Eg1S YTE)

SEQ ID NO: 224: Amino acid sequence of chimeric A-E57K_R58E-Fc (Eg1S YTE)

SEQ ID NO: 225: base sequence of chimeric A-E57R_R58D-Fc (Eg1S YTE)

SEQ ID NO: 226: Amino acid sequence of chimeric A-E57R_R58D-Fc (Eg1S YTE)

SEQ ID NO: 227: base sequence of chimeric A-E57K-Fc (Eg1S N434A)

SEQ ID NO: 228: Amino acid sequence of chimeric A-E57K-Fc (Eg1S N434A)

SEQ ID NO: 229: base sequence of chimeric A-E57L-Fc (Eg1S N434A)

SEQ ID NO: 230: Amino acid sequence of chimeric A-E57L-Fc (Eg1S N434A)

SEQ ID NO: 231: base sequence of chimeric A-E57R-Fc (Eg1S N434A)

SEQ ID NO: 232: Amino acid sequence of chimeric A-E57R-Fc (Eg1S N434A)

SEQ ID NO: 233: base sequence of chimeric A-E57V-Fc (Eg1S N434A)

SEQ ID NO: 234: Amino acid sequence of chimeric A-E57V-Fc (Eg1S N434A)

SEQ ID NO: 235: base sequence of chimeric A-E57K_R58D-Fc (Eg1S N434A)

SEQ ID NO: 236: Amino acid sequence of chimeric A-E57K_R58D-Fc (Eg1S N434A)

SEQ ID NO: 237: base sequence of chimeric A-E57K_R58E-Fc (Eg1S N434A)

SEQ ID NO: 238: Amino acid sequence of chimeric A-E57K_R58E-Fc (Eg1S N434A)

SEQ ID NO: 239: base sequence of chimeric A-E57R_R58D-Fc (Eg1S N434A)

SEQ ID NO: 240: Amino acid sequence of chimeric A-E57R_R58D-Fc (Eg1S N434A)

SEQ ID NO: 241: base sequence of chimeric A-E57K-Fc (Eg1S LALAGA)

SEQ ID NO: 242: Amino acid sequence of chimeric A-E57K-Fc (Eg1S LALAGA)

SEQ ID NO: 243: base sequence of chimeric A-E57L-Fc (Eg1S LALAGA)

SEQ ID NO: 244: Amino acid sequence of chimeric A-E57L-Fc (Eg1S LALAGA)

SEQ ID NO: 245: base sequence of chimeric A-E57R-Fc (Eg1S LALAGA)

SEQ ID NO: 246: Amino acid sequence of chimeric A-E57R-Fc (Eg1S LALAGA)

SEQ ID NO: 247: base sequence of chimeric A-E57V-Fc (Eg1S LALAGA)

SEQ ID NO: 248: Amino acid sequence of chimeric A-E57V-Fc (Eg1S LALAGA)

SEQ ID NO: 249: base sequence of chimeric A-E57K_R58D-Fc (Eg1S LALAGA)

SEQ ID NO: 250: Amino acid sequence of chimeric A-E57K_R58D-Fc (Eg1S LALAGA)

SEQ ID NO: 251: base sequence of chimeric A-E57K_R58E-Fc (Eg1S LALAGA)

SEQ ID NO: 252: Amino acid sequence of chimeric A-E57K_R58E-Fc (EglS LALAGA)

SEQ ID NO: 253: base sequence of chimeric A-E57R_R58D-Fc (Eg1S LALAGA)

SEQ ID NO: 254: Amino acid sequence of chimeric A-E57R_R58D-Fc (EglS LALAGA)

SEQ ID NO: 255: base sequence of chimeric A-E57K-Fc (Eg1S LALAGANA)

SEQ ID NO: 256: Amino acid sequence of chimeric A-E57K-Fc (Eg1S LALAGANA)

SEQ ID NO: 257: base sequence of chimeric A-E57L-Fc (Eg1S LALAGANA)

SEQ ID NO: 258: Amino acid sequence of chimeric A-E57L-Fc (Eg1S LALAGANA)

SEQ ID NO: 259: base sequence of chimeric A-E57R-Fc (Eg1S LALAGANA)

SEQ ID NO: 260: Amino acid sequence of chimeric A-E57R-Fc (Eg1S LALAGANA)

SEQ ID NO: 261: base sequence of chimeric A-E57V-Fc (Eg1S LALAGANA)

SEQ ID NO: 262: Amino acid sequence of chimeric A-E57V-Fc (Eg1S LALAGANA)

SEQ ID NO: 263: base sequence of chimeric A-E57K_R58D-Fc (Eg1S LALAGANA)

SEQ ID NO: 264: Amino acid sequence of chimeric A-E57K_R58D-Fc (Eg1S LALAGANA)

SEQ ID NO: 265: base sequence of chimeric A-E57K_R58E-Fc (Eg1S LALAGANA)

SEQ ID NO: 266: Amino acid sequence of chimeric A-E57K_R58E-Fc (Eg1S LALAGANA)

SEQ ID NO: 267: base sequence of chimeric A-E57R_R58D-Fc (Eg1S LALAGANA)

SEQ ID NO: 268: Amino acid sequence of chimeric A-E57R_R58D-Fc (Eg1S LALAGANA)

SEQ ID NO: 269: base sequence of chimeric A-E57A

SEQ ID NO: 270: Amino acid sequence of chimeric A-E57A

SEQ ID NO: 271: base sequence of chimeric A-E57F

SEQ ID NO: 272: Amino acid sequence of chimeric A-E57F

SEQ ID NO: 273: base sequence of chimeric A-E57H

SEQ ID NO: 274: amino acid sequence of chimeric A-E57H

SEQ ID NO: 275: base sequence of chimeric A-E57I

SEQ ID NO: 276: Amino acid sequence of chimeric A-E57I

SEQ ID NO: 277: base sequence of chimeric A-E57M

SEQ ID NO: 278: Amino acid sequence of chimeric A-E57M

SEQ ID NO: 279: base sequence of chimeric A-E57K_R58T

SEQ ID NO: 280: Amino acid sequence of chimeric A-E57K_R58T

SEQ ID NO: 281: base sequence of chimeric A-E57L_R58E

SEQ ID NO: 282: Amino acid sequence of chimeric A-E57L_R58E

SEQ ID NO: 283: base sequence of chimeric A-E57V_R58T

SEQ ID NO: 284: Amino acid sequence of chimeric A-E57V_R58T

SEQ ID NO: 285: base sequence of chimeric A-E57V_R58E

SEQ ID NO: 286: Amino acid sequence of chimeric A-E57V_R58E

SEQ ID NO: 287: base sequence of chimeric A-E57A-Fc (g4PEK)

SEQ ID NO: 288: Amino acid sequence of chimeric A-E57A-Fc (g4PEK)

SEQ ID NO: 289: base sequence of chimeric A-E57F-Fc (g4PEK)

SEQ ID NO: 290: Amino acid sequence of chimeric A-E57F-Fc (g4PEK)

SEQ ID NO: 291: base sequence of chimeric A-E57H-Fc (g4PEK)

SEQ ID NO: 292: Amino acid sequence of chimeric A-E57H-Fc (g4PEK)

SEQ ID NO: 293: base sequence of chimeric A-E57I-Fc (g4PEK)

SEQ ID NO: 294: Amino acid sequence of chimeric A-E57I-Fc (g4PEK)

SEQ ID NO: 295: base sequence of chimeric A-E57M-Fc (g4PEK)

SEQ ID NO: 296: Amino acid sequence of chimeric A-E57M-Fc (g4PEK)

SEQ ID NO: 297: base sequence of chimeric A-E57K_R58T-Fc (g4PEK)

SEQ ID NO: 298: Amino acid sequence of chimeric A-E57K_R58T-Fc (g4PEK)

SEQ ID NO: 299: base sequence of chimeric A-E57L_R58E-Fc (g4PEK)

SEQ ID NO: 300: amino acid sequence of chimeric A-E57L_R58E-Fc (g4PEK)

SEQ ID NO: 301: base sequence of chimeric A-E57V_R58T-Fc (g4PEK)

SEQ ID NO: 302: Amino acid sequence of chimeric A-E57V_R58T-Fc (g4PEK)

SEQ ID NO: 303: base sequence of chimeric A-E57V_R58E-Fc (g4PEK)

SEQ ID NO: 304: Amino acid sequence of chimeric A-E57V_R58E-Fc (g4PEK)

SEQ ID NO: 305: base sequence of soluble recombinant FLAG tag adduct of LIGHT

SEQ ID NO: 306: Amino acid sequence of soluble recombinant FLAG tag adduct of LIGHT

SEQ ID NO: 307: A nucleotide sequence in which a FLAG tag is inserted at the N-terminus of TL1A

SEQ ID NO: 308: amino acid sequence with FLAG tag inserted at the N-terminus of TL1A

SEQ ID NO: 309: A nucleotide sequence in which a FLAG tag is inserted at the N-terminus of FasL

SEQ ID NO: 310: amino acid sequence with FLAG tag inserted at the N-terminus of FasL

SEQ ID NO: 311: amino acid sequence of Fc (glS YTE)

SEQ ID NO: 312: amino acid sequence of Fc (glS N434A)

SEQ ID NO: 313: amino acid sequence of Fc (glS LALAGANA)

SEQ ID NO: 314: Amino acid sequence of chimeric A-Fc (glS YTE)

SEQ ID NO: 315: Amino acid sequence of chimeric A-Fc (g1S N434A)

SEQ ID NO: 316: Amino acid sequence of chimeric A-Fc (g1S LALAGANA)

SEQ ID NO: 317: Amino acid sequence of chimeric A-E57K-Fc (glS YTE)

SEQ ID NO: 318: Amino acid sequence of chimeric A-E57L-Fc (g1S YTE)

SEQ ID NO: 319: Amino acid sequence of chimeric A-E57R-Fc (glS YTE)

SEQ ID NO: 320: Amino acid sequence of chimeric A-E57V-Fc (g1S YTE)

SEQ ID NO: 321: Amino acid sequence of chimeric A-E57K_R58D-Fc (glS YTE)

SEQ ID NO: 322: Amino acid sequence of chimeric A-E57K_R58E-Fc (glS YTE)

SEQ ID NO: 323: Amino acid sequence of chimeric A-E57R_R58D-Fc (glS YTE)

SEQ ID NO: 324: Amino acid sequence of chimeric A-E57K-Fc (g1S N434A)

SEQ ID NO: 325: Amino acid sequence of chimeric A-E57L-Fc (g1S N434A)

SEQ ID NO: 326: Amino acid sequence of chimeric A-E57R-Fc (g1S N434A)

SEQ ID NO: 327: Amino acid sequence of chimeric A-E57V-Fc (g1S N434A)

SEQ ID NO: 328: Amino acid sequence of chimeric A-E57K_R58D-Fc (g1S N434A)

SEQ ID NO: 329: Amino acid sequence of chimeric A-E57K_R58E-Fc (g1S N434A)

SEQ ID NO: 330: Amino acid sequence of chimeric A-E57R_R58D-Fc (g1S N434A)

SEQ ID NO: 331: Amino acid sequence of chimeric A-E57K-Fc (g1S LALAGANA)

SEQ ID NO: 332: Amino acid sequence of chimeric A-E57L-Fc (g1S LALAGANA)

SEQ ID NO: 333: Amino acid sequence of chimeric A-E57R-Fc (g1S LALAGANA)

SEQ ID NO: 334: Amino acid sequence of chimeric A-E57V-Fc (g1S LALAGANA)

SEQ ID NO: 335: Amino acid sequence of chimeric A-E57K_R58D-Fc (g1S LALAGANA)

SEQ ID NO: 336: Amino acid sequence of chimeric A-E57K_R58E-Fc (glS LALAGANA)

SEQ ID NO: 337: Amino acid sequence of chimeric A-E57R_R58D-Fc (glS LALAGANA)

SEQ ID NO: 338: base sequence of Fc (IEGRMD g1S)

SEQ ID NO: 339: amino acid sequence of Fc (IEGRMD g1S)

SEQ ID NO: 340: amino acid sequence of R218Q-Fc

SEQ ID NO: 341: base sequence of FLAG-cynoLIGHT

SEQ ID NO: 342: amino acid sequence of FLAG-cynoLIGHT

SEQ ID NO: 343: A nucleotide sequence in which a cleavage site and an intracellular proline-rich region are deleted from FasL, a human DcR3 ligand, and a FLAG tag is inserted at the N-terminus

SEQ ID NO: 344: Amino acid sequence in which a cleavage site and an intracellular proline-rich region are deleted from FasL, a human DcR3 ligand, and a FLAG tag is inserted at the N-terminus

SEQ ID NO: 345: S195-His6 nucleotide sequence

SEQ ID NO: 346: amino acid sequence of S195-His6

SEQ ID NO: 347: base sequence of chimeric A-His6

SEQ ID NO: 348: Amino acid sequence of chimeric A-His6

SEQ ID NO: 349: base sequence of chimeric A-E57K-His6

SEQ ID NO: 350: amino acid sequence of chimeric A-E57K-His6

SEQ ID NO: 351: base sequence of chimeric A-E57K_R58E-His6

SEQ ID NO: 352: Amino acid sequence of chimeric A-E57K_R58E-His6

SEQ ID NO: 353: Optimized base sequence of chimera A

SEQ ID NO: 354: Optimized base sequence of chimeric A-E57K

SEQ ID NO: 355: Optimized base sequence of chimeric A-E57K_R58E

SEQUENCE LISTING <110> KYOWA KIRIN CO., LTD. <120> DcR3 variant <130> 233864PX <160> 355 <170> PatentIn version 3.5 <210> 1 <211> 900 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(900) <220> <223> Human DcR3 cDNA (Accession number: NM_003823.3) <400> 1 atg agg gcg ctg gag ggg cca ggc ctg tcg ctg ctg tgc ctg gtg ttg 48 Met Arg Ala Leu Glu Gly Pro Gly Leu Ser Leu Leu Cys Leu Val Leu 1 5 10 15 gcg ctg cct gcc ctg ctg ccg gtg ccg gct gta cgc gga gtg gca gaa 96 Ala Leu Pro Ala Leu Leu Pro Val Pro Ala Val Arg Gly Val Ala Glu 20 25 30 aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag cgg ctg gtg 144 Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu Arg Leu Val 35 40 45 tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg tgc cgc cga 192 Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro Cys Arg Arg 50 55 60 gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac tac acg cag 240 Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His Tyr Thr Gln 65 70 75 80 ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc ctc tgc ggg 288 Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val Leu Cys Gly 85 90 95 gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac aac cgt gcc 336 Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His Asn Arg Ala 100 105 110 tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc tgc ttg gag 384 Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe Cys Leu Glu 115 120 125 cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg ggc acc ccc 432 His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro Gly Thr Pro 130 135 140 agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc ttc tca gcc 480 Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr Phe Ser Ala 145 150 155 160 agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac tgc acg gcc 528 Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn Cys Thr Ala 165 170 175 ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat gac acc ctg 576 Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His Asp Thr Leu 180 185 190 tgc acc agc tgc act ggc ttc ccc ctc agc acc agg gta cca gga gct 624 Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val Pro Gly Ala 195 200 205 gag gag tgt gag cgt gcc gtc atc gac ttt gtg gct ttc cag gac atc 672 Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe Gln Asp Ile 210 215 220 tcc atc aag agg ctg cag cgg ctg ctg cag gcc ctc gag gcc ccg gag 720 Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu 225 230 235 240 ggc tgg ggt ccg aca cca agg gcg ggc cgc gcg gcc ttg cag ctg aag 768 Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys 245 250 255 ctg cgt cgg cgg ctc acg gag ctc ctg ggg gcg cag gac ggg gcg ctg 816 Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu 260 265 270 ctg gtg cgg ctg ctg cag gcg ctg cgc gtg gcc agg atg ccc ggg ctg 864 Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met Pro Gly Leu 275 280 285 gag cgg agc gtc cgt gag cgc ttc ctc cct gtg cac 900 Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 290 295 300 <210> 2 <211> 300 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of Human DcR3 (Accession number: NP_003814.1) <400> 2 Met Arg Ala Leu Glu Gly Pro Gly Leu Ser Leu Leu Cys Leu Val Leu 1 5 10 15 Ala Leu Pro Ala Leu Leu Pro Val Pro Ala Val Arg Gly Val Ala Glu 20 25 30 Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu Arg Leu Val 35 40 45 Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro Cys Arg Arg 50 55 60 Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His Tyr Thr Gln 65 70 75 80 Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val Leu Cys Gly 85 90 95 Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His Asn Arg Ala 100 105 110 Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe Cys Leu Glu 115 120 125 His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro Gly Thr Pro 130 135 140 Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr Phe Ser Ala 145 150 155 160 Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn Cys Thr Ala 165 170 175 Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His Asp Thr Leu 180 185 190 Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val Pro Gly Ala 195 200 205 Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe Gln Asp Ile 210 215 220 Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu 225 230 235 240 Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys 245 250 255 Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu 260 265 270 Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met Pro Gly Leu 275 280 285 Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 290 295 300 <210> 3 <211> 813 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(813) <220> <223> Nucleotide sequence of Human DcR3 (without a signal peptide) <400> 3 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac acg cag ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctc tgc ggg gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aac cgt gcc tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ttg gag cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggc acc ccc agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttc tca gcc agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gac acc ctg tgc acc agc tgc act ggc ttc ccc ctc agc acc agg gta 528 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 cca gga gct gag gag tgt gag cgt gcc gtc atc gac ttt gtg gct ttc 576 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 cag gac atc tcc atc aag agg ctg cag cgg ctg ctg cag gcc ctc gag 624 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 gcc ccg gag ggc tgg ggt ccg aca cca agg gcg ggc cgc gcg gcc ttg 672 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 cag ctg aag ctg cgt cgg cgg ctc acg gag ctc ctg ggg gcg cag gac 720 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 ggg gcg ctg ctg gtg cgg ctg ctg cag gcg ctg cgc gtg gcc agg atg 768 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 ccc ggg ctg gag cgg agc gtc cgt gag cgc ttc ctc cct gtg cac 813 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 260 265 270 <210> 4 <211> 271 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of Human DcR3 (without a signal peptide) <400> 4 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 260 265 270 <210> 5 <211> 123 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(123) <220> <223> Nucleotide sequence of CRD1 of Human DcR3 <400> 5 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt 123 Cys Arg Arg Asp Ser Pro Thr Thr Cys 35 40 <210> 6 <211> 41 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD1 of Human DcR3 <400> 6 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys 35 40 <210> 7 <211> 123 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(123) <220> <223> Nucleotide sequence of CRD2 of Human DcR3 <400> 7 tgt cca ccg cgc cac tac acg cag ttc tgg aac tac cta gag cgc tgc 48 Cys Pro Pro Arg His Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys 1 5 10 15 cgc tac tgc aac gtc ctc tgc ggg gag cgt gag gag gag gca cgg gct 96 Arg Tyr Cys Asn Val Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala 20 25 30 tgc cac gcc acc cac aac cgt gcc tgc 123 Cys His Ala Thr His Asn Arg Ala Cys 35 40 <210> 8 <211> 41 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD2 of Human DcR3 <400> 8 Cys Pro Pro Arg His Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys 1 5 10 15 Arg Tyr Cys Asn Val Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala 20 25 30 Cys His Ala Thr His Asn Arg Ala Cys 35 40 <210> 9 <211> 108 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(108) <220> <223> Nucleotide sequence of CRD3 of Human DcR3 <400> 9 tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc tgc ttg gag cac gca 48 Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe Cys Leu Glu His Ala 1 5 10 15 tcg tgt cca cct ggt gcc ggc gtg att gcc ccg ggc acc ccc agc cag 96 Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro Gly Thr Pro Ser Gln 20 25 30 aac acg cag tgc 108 Asn Thr Gln Cys 35 <210> 10 <211> 36 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD3 of Human DcR3 <400> 10 Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe Cys Leu Glu His Ala 1 5 10 15 Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro Gly Thr Pro Ser Gln 20 25 30 Asn Thr Gln Cys 35 <210> 11 <211> 123 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(123) <220> <223> Nucleotide sequence of CRD4 of Human DcR3 <400> 11 tgc ccc cca ggc acc ttc tca gcc agc agc tcc agc tca gag cag tgc 48 Cys Pro Pro Gly Thr Phe Ser Ala Ser Ser Ser Ser Ser Glu Gln Cys 1 5 10 15 cag ccc cac cgc aac tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca 96 Gln Pro His Arg Asn Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro 20 25 30 ggc tct tcc tcc cat gac acc ctg tgc 123 Gly Ser Ser Ser His Asp Thr Leu Cys 35 40 <210> 12 <211> 41 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD4 of Human DcR3 <400> 12 Cys Pro Pro Gly Thr Phe Ser Ala Ser Ser Ser Ser Ser Glu Gln Cys 1 5 10 15 Gln Pro His Arg Asn Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro 20 25 30 Gly Ser Ser Ser His Asp Thr Leu Cys 35 40 <210> 13 <211> 1203 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(1203) <220> <223> Nucleotide sequence of Human OPG (Accession number: NM_002546.3) <400> 13 atg aac aac ttg ctg tgc tgc gcg ctc gtg ttt ctg gac atc tcc att 48 Met Asn Asn Leu Leu Cys Cys Ala Leu Val Phe Leu Asp Ile Ser Ile 1 5 10 15 aag tgg acc acc cag gaa acg ttt cct cca aag tac ctt cat tat gac 96 Lys Trp Thr Thr Gln Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp 20 25 30 gaa gaa acc tct cat cag ctg ttg tgt gac aaa tgt cct cct ggt acc 144 Glu Glu Thr Ser His Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr 35 40 45 tac cta aaa caa cac tgt aca gca aag tgg aag acc gtg tgc gcc cct 192 Tyr Leu Lys Gln His Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro 50 55 60 tgc cct gac cac tac tac aca gac agc tgg cac acc agt gac gag tgt 240 Cys Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys 65 70 75 80 cta tac tgc agc ccc gtg tgc aag gag ctg cag tac gtc aag cag gag 288 Leu Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu 85 90 95 tgc aat cgc acc cac aac cgc gtg tgc gaa tgc aag gaa ggg cgc tac 336 Cys Asn Arg Thr His Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr 100 105 110 ctt gag ata gag ttc tgc ttg aaa cat agg agc tgc cct cct gga ttt 384 Leu Glu Ile Glu Phe Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe 115 120 125 gga gtg gtg caa gct gga acc cca gag cga aat aca gtt tgc aaa aga 432 Gly Val Val Gln Ala Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg 130 135 140 tgt cca gat ggg ttc ttc tca aat gag acg tca tct aaa gca ccc tgt 480 Cys Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys 145 150 155 160 aga aaa cac aca aat tgc agt gtc ttt ggt ctc ctg cta act cag aaa 528 Arg Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys 165 170 175 gga aat gca aca cac gac aac ata tgt tcc gga aac agt gaa tca act 576 Gly Asn Ala Thr His Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr 180 185 190 caa aaa tgt gga ata gat gtt acc ctg tgt gag gag gca ttc ttc agg 624 Gln Lys Cys Gly Ile Asp Val Thr Leu Cys Glu Glu Ala Phe Phe Arg 195 200 205 ttt gct gtt cct aca aag ttt acg cct aac tgg ctt agt gtc ttg gta 672 Phe Ala Val Pro Thr Lys Phe Thr Pro Asn Trp Leu Ser Val Leu Val 210 215 220 gac aat ttg cct ggc acc aaa gta aac gca gag agt gta gag agg ata 720 Asp Asn Leu Pro Gly Thr Lys Val Asn Ala Glu Ser Val Glu Arg Ile 225 230 235 240 aaa cgg caa cac agc tca caa gaa cag act ttc cag ctg ctg aag tta 768 Lys Arg Gln His Ser Ser Gln Glu Gln Thr Phe Gln Leu Leu Lys Leu 245 250 255 tgg aaa cat caa aac aaa gac caa gat ata gtc aag aag atc atc caa 816 Trp Lys His Gln Asn Lys Asp Gln Asp Ile Val Lys Lys Ile Ile Gln 260 265 270 gat att gac ctc tgt gaa aac agc gtg cag cgg cac att gga cat gct 864 Asp Ile Asp Leu Cys Glu Asn Ser Val Gln Arg His Ile Gly His Ala 275 280 285 aac ctc acc ttc gag cag ctt cgt agc ttg atg gaa agc tta ccg gga 912 Asn Leu Thr Phe Glu Gln Leu Arg Ser Leu Met Glu Ser Leu Pro Gly 290 295 300 aag aaa gtg gga gca gaa gac att gaa aaa aca ata aag gca tgc aaa 960 Lys Lys Val Gly Ala Glu Asp Ile Glu Lys Thr Ile Lys Ala Cys Lys 305 310 315 320 ccc agt gac cag atc ctg aag ctg ctc agt ttg tgg cga ata aaa aat 1008 Pro Ser Asp Gln Ile Leu Lys Leu Leu Ser Leu Trp Arg Ile Lys Asn 325 330 335 ggc gac caa gac acc ttg aag ggc cta atg cac gca cta aag cac tca 1056 Gly Asp Gln Asp Thr Leu Lys Gly Leu Met His Ala Leu Lys His Ser 340 345 350 aag acg tac cac ttt ccc aaa act gtc act cag agt cta aag aag acc 1104 Lys Thr Tyr His Phe Pro Lys Thr Val Thr Gln Ser Leu Lys Lys Thr 355 360 365 atc agg ttc ctt cac agc ttc aca atg tac aaa ttg tat cag aag tta 1152 Ile Arg Phe Leu His Ser Phe Thr Met Tyr Lys Leu Tyr Gln Lys Leu 370 375 380 ttt tta gaa atg ata ggt aac cag gtc caa tca gta aaa ata agc tgc 1200 Phe Leu Glu Met Ile Gly Asn Gln Val Gln Ser Val Lys Ile Ser Cys 385 390 395 400 tta 1203 Leu <210> 14 <211> 401 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of Human OPG (Accession number: NP_002537.3) <400> 14 Met Asn Asn Leu Leu Cys Cys Ala Leu Val Phe Leu Asp Ile Ser Ile 1 5 10 15 Lys Trp Thr Thr Gln Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp 20 25 30 Glu Glu Thr Ser His Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr 35 40 45 Tyr Leu Lys Gln His Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro 50 55 60 Cys Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys 65 70 75 80 Leu Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu 85 90 95 Cys Asn Arg Thr His Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr 100 105 110 Leu Glu Ile Glu Phe Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe 115 120 125 Gly Val Val Gln Ala Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg 130 135 140 Cys Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys 145 150 155 160 Arg Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys 165 170 175 Gly Asn Ala Thr His Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr 180 185 190 Gln Lys Cys Gly Ile Asp Val Thr Leu Cys Glu Glu Ala Phe Phe Arg 195 200 205 Phe Ala Val Pro Thr Lys Phe Thr Pro Asn Trp Leu Ser Val Leu Val 210 215 220 Asp Asn Leu Pro Gly Thr Lys Val Asn Ala Glu Ser Val Glu Arg Ile 225 230 235 240 Lys Arg Gln His Ser Ser Gln Glu Gln Thr Phe Gln Leu Leu Lys Leu 245 250 255 Trp Lys His Gln Asn Lys Asp Gln Asp Ile Val Lys Lys Ile Ile Gln 260 265 270 Asp Ile Asp Leu Cys Glu Asn Ser Val Gln Arg His Ile Gly His Ala 275 280 285 Asn Leu Thr Phe Glu Gln Leu Arg Ser Leu Met Glu Ser Leu Pro Gly 290 295 300 Lys Lys Val Gly Ala Glu Asp Ile Glu Lys Thr Ile Lys Ala Cys Lys 305 310 315 320 Pro Ser Asp Gln Ile Leu Lys Leu Leu Ser Leu Trp Arg Ile Lys Asn 325 330 335 Gly Asp Gln Asp Thr Leu Lys Gly Leu Met His Ala Leu Lys His Ser 340 345 350 Lys Thr Tyr His Phe Pro Lys Thr Val Thr Gln Ser Leu Lys Lys Thr 355 360 365 Ile Arg Phe Leu His Ser Phe Thr Met Tyr Lys Leu Tyr Gln Lys Leu 370 375 380 Phe Leu Glu Met Ile Gly Asn Gln Val Gln Ser Val Lys Ile Ser Cys 385 390 395 400 Leu <210> 15 <211> 1140 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(1140) <220> <223> Nucleotide sequence of Human OPG (without a signal peptide) <400> 15 gaa acg ttt cct cca aag tac ctt cat tat gac gaa gaa acc tct cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ttg tgt gac aaa tgt cct cct ggt acc tac cta aaa caa cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt aca gca aag tgg aag acc gtg tgc gcc cct tgc cct gac cac tac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Asp His Tyr 35 40 45 tac aca gac agc tgg cac acc agt gac gag tgt cta tac tgc agc ccc 192 Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro 50 55 60 gtg tgc aag gag ctg cag tac gtc aag cag gag tgc aat cgc acc cac 240 Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His 65 70 75 80 aac cgc gtg tgc gaa tgc aag gaa ggg cgc tac ctt gag ata gag ttc 288 Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe 85 90 95 tgc ttg aaa cat agg agc tgc cct cct gga ttt gga gtg gtg caa gct 336 Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 gga acc cca gag cga aat aca gtt tgc aaa aga tgt cca gat ggg ttc 384 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 ttc tca aat gag acg tca tct aaa gca ccc tgt aga aaa cac aca aat 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgc agt gtc ttt ggt ctc ctg cta act cag aaa gga aat gca aca cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac ata tgt tcc gga aac agt gaa tca act caa aaa tgt gga ata 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Gly Ile 165 170 175 gat gtt acc ctg tgt gag gag gca ttc ttc agg ttt gct gtt cct aca 576 Asp Val Thr Leu Cys Glu Glu Ala Phe Phe Arg Phe Ala Val Pro Thr 180 185 190 aag ttt acg cct aac tgg ctt agt gtc ttg gta gac aat ttg cct ggc 624 Lys Phe Thr Pro Asn Trp Leu Ser Val Leu Val Asp Asn Leu Pro Gly 195 200 205 acc aaa gta aac gca gag agt gta gag agg ata aaa cgg caa cac agc 672 Thr Lys Val Asn Ala Glu Ser Val Glu Arg Ile Lys Arg Gln His Ser 210 215 220 tca caa gaa cag act ttc cag ctg ctg aag tta tgg aaa cat caa aac 720 Ser Gln Glu Gln Thr Phe Gln Leu Leu Lys Leu Trp Lys His Gln Asn 225 230 235 240 aaa gac caa gat ata gtc aag aag atc atc caa gat att gac ctc tgt 768 Lys Asp Gln Asp Ile Val Lys Lys Ile Ile Gln Asp Ile Asp Leu Cys 245 250 255 gaa aac agc gtg cag cgg cac att gga cat gct aac ctc acc ttc gag 816 Glu Asn Ser Val Gln Arg His Ile Gly His Ala Asn Leu Thr Phe Glu 260 265 270 cag ctt cgt agc ttg atg gaa agc tta ccg gga aag aaa gtg gga gca 864 Gln Leu Arg Ser Leu Met Glu Ser Leu Pro Gly Lys Lys Val Gly Ala 275 280 285 gaa gac att gaa aaa aca ata aag gca tgc aaa ccc agt gac cag atc 912 Glu Asp Ile Glu Lys Thr Ile Lys Ala Cys Lys Pro Ser Asp Gln Ile 290 295 300 ctg aag ctg ctc agt ttg tgg cga ata aaa aat ggc gac caa gac acc 960 Leu Lys Leu Leu Ser Leu Trp Arg Ile Lys Asn Gly Asp Gln Asp Thr 305 310 315 320 ttg aag ggc cta atg cac gca cta aag cac tca aag acg tac cac ttt 1008 Leu Lys Gly Leu Met His Ala Leu Lys His Ser Lys Thr Tyr His Phe 325 330 335 ccc aaa act gtc act cag agt cta aag aag acc atc agg ttc ctt cac 1056 Pro Lys Thr Val Thr Gln Ser Leu Lys Lys Thr Ile Arg Phe Leu His 340 345 350 agc ttc aca atg tac aaa ttg tat cag aag tta ttt tta gaa atg ata 1104 Ser Phe Thr Met Tyr Lys Leu Tyr Gln Lys Leu Phe Leu Glu Met Ile 355 360 365 ggt aac cag gtc caa tca gta aaa ata agc tgc tta 1140 Gly Asn Gln Val Gln Ser Val Lys Ile Ser Cys Leu 370 375 380 <210> 16 <211> 380 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of Human OPG (without a signal peptide) <400> 16 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Asp His Tyr 35 40 45 Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys Leu Tyr Cys Ser Pro 50 55 60 Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu Cys Asn Arg Thr His 65 70 75 80 Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe 85 90 95 Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Gly Ile 165 170 175 Asp Val Thr Leu Cys Glu Glu Ala Phe Phe Arg Phe Ala Val Pro Thr 180 185 190 Lys Phe Thr Pro Asn Trp Leu Ser Val Leu Val Asp Asn Leu Pro Gly 195 200 205 Thr Lys Val Asn Ala Glu Ser Val Glu Arg Ile Lys Arg Gln His Ser 210 215 220 Ser Gln Glu Gln Thr Phe Gln Leu Leu Lys Leu Trp Lys His Gln Asn 225 230 235 240 Lys Asp Gln Asp Ile Val Lys Lys Ile Ile Gln Asp Ile Asp Leu Cys 245 250 255 Glu Asn Ser Val Gln Arg His Ile Gly His Ala Asn Leu Thr Phe Glu 260 265 270 Gln Leu Arg Ser Leu Met Glu Ser Leu Pro Gly Lys Lys Val Gly Ala 275 280 285 Glu Asp Ile Glu Lys Thr Ile Lys Ala Cys Lys Pro Ser Asp Gln Ile 290 295 300 Leu Lys Leu Leu Ser Leu Trp Arg Ile Lys Asn Gly Asp Gln Asp Thr 305 310 315 320 Leu Lys Gly Leu Met His Ala Leu Lys His Ser Lys Thr Tyr His Phe 325 330 335 Pro Lys Thr Val Thr Gln Ser Leu Lys Lys Thr Ile Arg Phe Leu His 340 345 350 Ser Phe Thr Met Tyr Lys Leu Tyr Gln Lys Leu Phe Leu Glu Met Ile 355 360 365 Gly Asn Gln Val Gln Ser Val Lys Ile Ser Cys Leu 370 375 380 <210> 17 <211> 123 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(123) <220> <223> Nucleotide sequence of CRD1 of Human OPG <400> 17 gaa acg ttt cct cca aag tac ctt cat tat gac gaa gaa acc tct cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ttg tgt gac aaa tgt cct cct ggt acc tac cta aaa caa cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt aca gca aag tgg aag acc gtg tgc 123 Cys Thr Ala Lys Trp Lys Thr Val Cys 35 40 <210> 18 <211> 41 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD1 of Human OPG <400> 18 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys 35 40 <210> 19 <211> 123 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(123) <220> <223> Nucleotide sequence of CRD2 of Human OPG <400> 19 tgc cct gac cac tac tac aca gac agc tgg cac acc agt gac gag tgt 48 Cys Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys 1 5 10 15 cta tac tgc agc ccc gtg tgc aag gag ctg cag tac gtc aag cag gag 96 Leu Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu 20 25 30 tgc aat cgc acc cac aac cgc gtg tgc 123 Cys Asn Arg Thr His Asn Arg Val Cys 35 40 <210> 20 <211> 41 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD2 of Human OPG <400> 20 Cys Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys 1 5 10 15 Leu Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu 20 25 30 Cys Asn Arg Thr His Asn Arg Val Cys 35 40 <210> 21 <211> 108 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(108) <220> <223> Nucleotide sequence of CRD3 of Human OPG <400> 21 tgc aag gaa ggg cgc tac ctt gag ata gag ttc tgc ttg aaa cat agg 48 Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe Cys Leu Lys His Arg 1 5 10 15 agc tgc cct cct gga ttt gga gtg gtg caa gct gga acc cca gag cga 96 Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala Gly Thr Pro Glu Arg 20 25 30 aat aca gtt tgc 108 Asn Thr Val Cys 35 <210> 22 <211> 36 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD3 of Human OPG <400> 22 Cys Lys Glu Gly Arg Tyr Leu Glu Ile Glu Phe Cys Leu Lys His Arg 1 5 10 15 Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala Gly Thr Pro Glu Arg 20 25 30 Asn Thr Val Cys 35 <210> 23 <211> 123 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(123) <220> <223> Nucleotide sequence of CRD4 of Human OPG <400> 23 tgt cca gat ggg ttc ttc tca aat gag acg tca tct aaa gca ccc tgt 48 Cys Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys 1 5 10 15 aga aaa cac aca aat tgc agt gtc ttt ggt ctc ctg cta act cag aaa 96 Arg Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys 20 25 30 gga aat gca aca cac gac aac ata tgt 123 Gly Asn Ala Thr His Asp Asn Ile Cys 35 40 <210> 24 <211> 41 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of CRD4 of Human OPG <400> 24 Cys Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys 1 5 10 15 Arg Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys 20 25 30 Gly Asn Ala Thr His Asp Asn Ile Cys 35 40 <210> 25 <211> 813 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(813) <220> <223> Nucleotide sequence of Chimera B-HBD <400> 25 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct ccc ggc act 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttt tct gcc tct tcc agt tca tca gag caa tgc cag cct cac aga aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgt acc gct ctg ggg ctc gcc ctg aat gtc ccc ggc tct tca agt cac 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gat acc ctg tgc acc agc tgc act ggc ttc ccc ctc agc acc agg gta 528 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 cca gga gct gag gag tgt gag cgt gcc gtc atc gac ttt gtg gct ttc 576 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 cag gac atc tcc atc aag agg ctg cag cgg ctg ctg cag gcc ctc gag 624 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 gcc ccg gag ggc tgg ggt ccg aca cca agg gcg ggc cgc gcg gcc ttg 672 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 cag ctg aag ctg cgt cgg cgg ctc acg gag ctc ctg ggg gcg cag gac 720 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 ggg gcg ctg ctg gtg cgg ctg ctg cag gcg ctg cgc gtg gcc agg atg 768 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 ccc ggg ctg gag cgg agc gtc cgt gag cgc ttc ctc cct gtg cac 813 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 260 265 270 <210> 26 <211> 271 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera B-HBD <400> 26 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 260 265 270 <210> 27 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of Chimera C-HBD <400> 27 gta gca gag aca ccc acc tat cca tgg agg gac gct gag acc ggt gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctc gtt tgc gcc caa tgt cca cca ggg acc ttc gtt cag cgg ccc 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cga cgg gac agt cca aca aca tgt ggc cca tgc cca cca aga cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 28 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera C-HBD <400> 28 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 29 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of Chimera A-HBD <400> 29 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 30 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-HBD <400> 30 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 31 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of 103-123OPG-HBD <400> 31 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt cct cca gga ttt ggg gtg gtc caa gca 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 ggg aca ccc gag cgg aac acc gta tgc aaa cga tgt cct gac ggc ttt 384 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 32 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of 103-123OPG-HBD <400> 32 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 33 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of N131S/N144S-HBD <400> 33 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct agt gag act tca agt aaa gct ccc tgc cgc aag cac acc agc 432 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 34 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of N131S/N144S-HBD <400> 34 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 35 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of T133A/S146A-HBD <400> 35 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag gct tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt gct gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 36 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of T133A/S146A-HBD <400> 36 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 37 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of N131S/N144S/N157S-HBD <400> 37 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct agt gag act tca agt aaa gct ccc tgc cgc aag cac acc agc 432 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg agc gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Ser Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 38 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of N131S/N144S/N157S-HBD <400> 38 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Ser Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 39 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of T133A/S146A/T159A-HBD <400> 39 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag gct tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt gct gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca gct cac 480 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Ala His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 40 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of T133A/S146A/T159A-HBD <400> 40 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Ala His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 41 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of Chimera A-E57K-HBD <400> 41 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 42 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-HBD <400> 42 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 43 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of Chimera A-E57L-HBD <400> 43 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 44 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-HBD <400> 44 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 45 <211> 834 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(834) <220> <223> Nucleotide sequence of Chimera A-R60K-HBD <400> 45 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc aag tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Lys Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag tgc act ggc 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt gag cgt gcc 576 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag agg ctg cag 624 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt ccg aca cca 672 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg cgg ctc acg 720 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg ctg ctg cag 768 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc gtc cgt gag 816 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 cgc ttc ctc cct gtg cac 834 Arg Phe Leu Pro Val His 275 <210> 46 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-R60K-HBD <400> 46 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Lys Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Cys Thr Gly 165 170 175 Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys Glu Arg Ala 180 185 190 Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys Arg Leu Gln 195 200 205 Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly Pro Thr Pro 210 215 220 Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg Arg Leu Thr 225 230 235 240 Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg Leu Leu Gln 245 250 255 Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser Val Arg Glu 260 265 270 Arg Phe Leu Pro Val His 275 <210> 47 <211> 315 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(315) <220> <223> Nucleotide sequence of HBD of Wild type DcR3 <400> 47 tgc act ggc ttc ccc ctc agc acc agg gta cca gga gct gag gag tgt 48 Cys Thr Gly Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys 1 5 10 15 gag cgt gcc gtc atc gac ttt gtg gct ttc cag gac atc tcc atc aag 96 Glu Arg Ala Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys 20 25 30 agg ctg cag cgg ctg ctg cag gcc ctc gag gcc ccg gag ggc tgg ggt 144 Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly 35 40 45 ccg aca cca agg gcg ggc cgc gcg gcc ttg cag ctg aag ctg cgt cgg 192 Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg 50 55 60 cgg ctc acg gag ctc ctg ggg gcg cag gac ggg gcg ctg ctg gtg cgg 240 Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg 65 70 75 80 ctg ctg cag gcg ctg cgc gtg gcc agg atg ccc ggg ctg gag cgg agc 288 Leu Leu Gln Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser 85 90 95 gtc cgt gag cgc ttc ctc cct gtg cac 315 Val Arg Glu Arg Phe Leu Pro Val His 100 105 <210> 48 <211> 105 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of HBD of Wild type DcR3 <400> 48 Cys Thr Gly Phe Pro Leu Ser Thr Arg Val Pro Gly Ala Glu Glu Cys 1 5 10 15 Glu Arg Ala Val Ile Asp Phe Val Ala Phe Gln Asp Ile Ser Ile Lys 20 25 30 Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu Ala Pro Glu Gly Trp Gly 35 40 45 Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu Gln Leu Lys Leu Arg Arg 50 55 60 Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp Gly Ala Leu Leu Val Arg 65 70 75 80 Leu Leu Gln Ala Leu Arg Val Ala Arg Met Pro Gly Leu Glu Arg Ser 85 90 95 Val Arg Glu Arg Phe Leu Pro Val His 100 105 <210> 49 <211> 498 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(498) <220> <223> Nucleotide sequence of Chimera B <400> 49 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct ccc ggc act 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttt tct gcc tct tcc agt tca tca gag caa tgc cag cct cac aga aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgt acc gct ctg ggg ctc gcc ctg aat gtc ccc ggc tct tca agt cac 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gat acc ctg tgc acc agc 498 Asp Thr Leu Cys Thr Ser 165 <210> 50 <211> 166 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera B <400> 50 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser 165 <210> 51 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera C <400> 51 gta gca gag aca ccc acc tat cca tgg agg gac gct gag acc ggt gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctc gtt tgc gcc caa tgt cca cca ggg acc ttc gtt cag cgg ccc 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cga cgg gac agt cca aca aca tgt ggc cca tgc cca cca aga cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 52 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera C <400> 52 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 53 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A <400> 53 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 54 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A <400> 54 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 55 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of 103-123OPG <400> 55 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt cct cca gga ttt ggg gtg gtc caa gca 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 ggg aca ccc gag cgg aac acc gta tgc aaa cga tgt cct gac ggc ttt 384 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 56 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of 103-123OPG <400> 56 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 57 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of N131S/N144S <400> 57 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct agt gag act tca agt aaa gct ccc tgc cgc aag cac acc agc 432 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 58 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of N131S/N144S <400> 58 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 59 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of T133A/S146A <400> 59 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag gct tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt gct gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 60 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of T133A/S146A <400> 60 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 61 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of N131S/N144S/N157S <400> 61 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct agt gag act tca agt aaa gct ccc tgc cgc aag cac acc agc 432 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg agc gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Ser Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 62 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of N131S/N144S/N157S <400> 62 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Ser Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 63 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of T133A/S146A/T159A <400> 63 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag gct tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt gct gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca gct cac 480 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Ala His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 64 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of T133A/S146A/T159A <400> 64 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Ala His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 65 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57K <400> 65 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 66 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K <400> 66 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 67 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57L <400> 67 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 68 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L <400> 68 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 69 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-R60K <400> 69 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc aag tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Lys Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 70 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-R60K <400> 70 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Lys Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 71 <211> 693 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(693) <220> <223> Nucleotide sequence of Fc (g1S) <400> 71 ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct 48 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag 96 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro Lys 20 25 30 gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg 144 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac 192 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac 240 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac 288 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc 336 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga 384 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag 432 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac 480 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag 528 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc 576 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca 624 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc 672 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 ctc tcc ctg tct ccg ggt aaa 693 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 72 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (g1S) <400> 72 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 73 <211> 687 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(687) <220> <223> Nucleotide sequence of Fc (g4PEK) <400> 73 gag tcc aaa tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc 48 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe 1 5 10 15 gag ggg gga cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act 96 Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 ctc atg atc tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg 144 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 agc cag gaa gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg 192 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 65 70 75 80 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 288 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 85 90 95 aac ggc aag gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc 336 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 tcc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca 384 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 cag gtg tac acc ctg ccc cca tcc cag gag gag atg acc aag aac cag 432 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 130 135 140 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc 480 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 145 150 155 160 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 528 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta 576 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 180 185 190 acc gtg gac aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc 624 Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 195 200 205 gtg atg cat gag gct ctg cac aac cac tac aca cag aag agc ctc tcc 672 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 ctg tct ctg ggt aaa 687 Leu Ser Leu Gly Lys 225 <210> 74 <211> 229 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (g4PEK) <400> 74 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe 1 5 10 15 Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 65 70 75 80 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 85 90 95 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 130 135 140 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 145 150 155 160 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 180 185 190 Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 195 200 205 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 Leu Ser Leu Gly Lys 225 <210> 75 <211> 1209 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1209) <220> <223> Nucleotide sequence of Chimera B-Fc (IEGRMD-g1S) <400> 75 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct ccc ggc act 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttt tct gcc tct tcc agt tca tca gag caa tgc cag cct cac aga aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgt acc gct ctg ggg ctc gcc ctg aat gtc ccc ggc tct tca agt cac 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gat acc ctg tgc acc agc atc gag ggg cgc atg gat ccc aaa tct tcc 528 Asp Thr Leu Cys Thr Ser Ile Glu Gly Arg Met Asp Pro Lys Ser Ser 165 170 175 gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc ctg ggg 576 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 180 185 190 gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc ctc atg 624 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 195 200 205 atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg agc cac 672 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 210 215 220 gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag gtg 720 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 225 230 235 240 cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc acg tac 768 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 245 250 255 cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aat ggc 816 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 260 265 270 aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc ccc atc 864 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 275 280 285 gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca cag gtg 912 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 290 295 300 tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag gtc agc 960 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 305 310 315 320 ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc gtg gag 1008 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 325 330 335 tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc 1056 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 340 345 350 gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc acc gtg 1104 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 355 360 365 gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc gtg atg 1152 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 370 375 380 cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc ctg tct 1200 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 385 390 395 400 ccg ggt aaa 1209 Pro Gly Lys <210> 76 <211> 403 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera B-Fc (IEGRMD-g1S) <400> 76 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Ile Glu Gly Arg Met Asp Pro Lys Ser Ser 165 170 175 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 180 185 190 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 195 200 205 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 210 215 220 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 225 230 235 240 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 245 250 255 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 260 265 270 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 275 280 285 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 290 295 300 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 305 310 315 320 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 325 330 335 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 340 345 350 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 355 360 365 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 370 375 380 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 385 390 395 400 Pro Gly Lys <210> 77 <211> 1230 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1230) <220> <223> Nucleotide sequence of Chimera C-Fc (IEGRMD-g1S) <400> 77 gta gca gag aca ccc acc tat cca tgg agg gac gct gag acc ggt gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctc gtt tgc gcc caa tgt cca cca ggg acc ttc gtt cag cgg ccc 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cga cgg gac agt cca aca aca tgt ggc cca tgc cca cca aga cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag atc gag ggg 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Ile Glu Gly 165 170 175 cgc atg gat ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc 576 Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys 180 185 190 cca gca cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca 624 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 195 200 205 aaa ccc aag gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc 672 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 210 215 220 gtg gtg gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg 720 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 225 230 235 240 tac gtg gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag 768 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 245 250 255 gag cag tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg 816 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 260 265 270 cac cag gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac 864 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 275 280 285 aaa gcc ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg 912 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 290 295 300 cag ccc cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag 960 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 305 310 315 320 ctg acc aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat 1008 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 325 330 335 ccc agc gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac 1056 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 340 345 350 aac tac aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc 1104 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 355 360 365 ctc tac agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac 1152 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 370 375 380 gtc ttc tca tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg 1200 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 385 390 395 400 cag aag agc ctc tcc ctg tct ccg ggt aaa 1230 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 405 410 <210> 78 <211> 410 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera C-Fc (IEGRMD-g1S) <400> 78 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Ile Glu Gly 165 170 175 Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys 180 185 190 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 195 200 205 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 210 215 220 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 225 230 235 240 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 245 250 255 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 260 265 270 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 275 280 285 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 290 295 300 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 305 310 315 320 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 325 330 335 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 340 345 350 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 355 360 365 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 370 375 380 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 385 390 395 400 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 405 410 <210> 79 <211> 1230 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1230) <220> <223> Nucleotide sequence of Chimera A-Fc (IEGRMD-g1S) <400> 79 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag atc gag ggg 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Ile Glu Gly 165 170 175 cgc atg gat ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc 576 Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys 180 185 190 cca gca cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca 624 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 195 200 205 aaa ccc aag gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc 672 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 210 215 220 gtg gtg gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg 720 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 225 230 235 240 tac gtg gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag 768 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 245 250 255 gag cag tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg 816 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 260 265 270 cac cag gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac 864 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 275 280 285 aaa gcc ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg 912 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 290 295 300 cag ccc cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag 960 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 305 310 315 320 ctg acc aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat 1008 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 325 330 335 ccc agc gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac 1056 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 340 345 350 aac tac aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc 1104 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 355 360 365 ctc tac agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac 1152 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 370 375 380 gtc ttc tca tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg 1200 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 385 390 395 400 cag aag agc ctc tcc ctg tct ccg ggt aaa 1230 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 405 410 <210> 80 <211> 410 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (IEGRMD-g1S) <400> 80 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Ile Glu Gly 165 170 175 Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys 180 185 190 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 195 200 205 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 210 215 220 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 225 230 235 240 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 245 250 255 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 260 265 270 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 275 280 285 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 290 295 300 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 305 310 315 320 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 325 330 335 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 340 345 350 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 355 360 365 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 370 375 380 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 385 390 395 400 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 405 410 <210> 81 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-Fc (g4PEK) <400> 81 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 82 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (g4PEK) <400> 82 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 83 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of 103-123OPG-Fc (g4PEK) <400> 83 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt cct cca gga ttt ggg gtg gtc caa gca 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 ggg aca ccc gag cgg aac acc gta tgc aaa cga tgt cct gac ggc ttt 384 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 84 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of 103-123OPG-Fc (g4PEK) <400> 84 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Phe Gly Val Val Gln Ala 100 105 110 Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 85 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of N131S/N144S-Fc (g4PEK) <400> 85 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct agt gag act tca agt aaa gct ccc tgc cgc aag cac acc agc 432 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 86 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of N131S/N144S-Fc (g4PEK) <400> 86 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 87 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of T133A/S146A-Fc (g4PEK) <400> 87 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag gct tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt gct gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 88 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of T133A/S146A-Fc (g4PEK) <400> 88 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 89 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of N131S/N144S/N157S-Fc (g4PEK) <400> 89 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct agt gag act tca agt aaa gct ccc tgc cgc aag cac acc agc 432 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg agc gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Ser Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 90 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of N131S/N144S/N157S-Fc (g4PEK) <400> 90 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Ser Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Ser 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Ser Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 91 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of T133A/S146A/T159A-Fc (g4PEK) <400> 91 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag gct tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt gct gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca gct cac 480 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Ala His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 92 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of T133A/S146A/T159A-Fc (g4PEK) <400> 92 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Ala Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ala Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Ala His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 93 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57K-Fc (g4PEK) <400> 93 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 94 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (g4PEK) <400> 94 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 95 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57L-Fc (g4PEK) <400> 95 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 96 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (g4PEK) <400> 96 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 97 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-R60K-Fc (g4PEK) <400> 97 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc aag tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Lys Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 98 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-R60K-Fc (g4PEK) <400> 98 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Lys Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 99 <211> 1524 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1524) <220> <223> Nucleotide sequence of DcR3 FL-Fc <400> 99 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac acg cag ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctc tgc ggg gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aac cgt gcc tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ttg gag cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggc acc ccc agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttc tca gcc agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gac acc ctg tgc acc agc tgc act ggc ttc ccc ctc agc acc agg gta 528 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 cca gga gct gag gag tgt gag cgt gcc gtc atc gac ttt gtg gct ttc 576 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 cag gac atc tcc atc aag agg ctg cag cgg ctg ctg cag gcc ctc gag 624 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 gcc ccg gag ggc tgg ggt ccg aca cca agg gcg ggc cgc gcg gcc ttg 672 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 cag ctg aag ctg cgt cgg cgg ctc acg gag ctc ctg ggg gcg cag gac 720 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 ggg gcg ctg ctg gtg cgg ctg ctg cag gcg ctg cgc gtg gcc agg atg 768 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 ccc ggg ctg gag cgg agc gtc cgt gag cgc ttc ctc cct gtg cac atc 816 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His Ile 260 265 270 gag ggg cgc atg gat ccc aaa tct tcc gac aaa act cac aca tgc cca 864 Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro 275 280 285 ccg tgc cca gca cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc 912 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 290 295 300 ccc cca aaa ccc aag gac acc ctc atg atc tcc cgg acc cct gag gtc 960 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 305 310 315 320 aca tgc gtg gtg gtg gac gtg agc cac gaa gac cct gag gtc aag ttc 1008 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 325 330 335 aac tgg tac gtg gac ggc gtg gag gtg cat aat gcc aag aca aag ccg 1056 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 340 345 350 cgg gag gag cag tac aac agc acg tac cgt gtg gtc agc gtc ctc acc 1104 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 355 360 365 gtc ctg cac cag gac tgg ctg aat ggc aag gag tac aag tgc aag gtc 1152 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 370 375 380 tcc aac aaa gcc ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc 1200 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 385 390 395 400 aaa ggg cag ccc cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg 1248 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 405 410 415 gat gag ctg acc aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc 1296 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 420 425 430 ttc tat ccc agc gac atc gcc gtg gag tgg gag agc aat ggg cag ccg 1344 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 435 440 445 gag aac aac tac aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc 1392 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 450 455 460 ttc ttc ctc tac agc aag ctc acc gtg gac aag agc agg tgg cag cag 1440 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 465 470 475 480 ggg aac gtc ttc tca tgc tcc gtg atg cat gag gct ctg cac aac cac 1488 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 485 490 495 tac acg cag aag agc ctc tcc ctg tct ccg ggt aaa 1524 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 500 505 <210> 100 <211> 508 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of DcR3 FL-Fc <400> 100 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His Ile 260 265 270 Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro 275 280 285 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 290 295 300 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 305 310 315 320 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 325 330 335 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 340 345 350 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 355 360 365 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 370 375 380 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 385 390 395 400 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 405 410 415 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 420 425 430 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 435 440 445 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 450 455 460 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 465 470 475 480 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 485 490 495 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 500 505 <210> 101 <211> 1209 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1209) <220> <223> Nucleotide sequence of S195-Fc <400> 101 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac acg cag ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctc tgc ggg gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aac cgt gcc tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ttg gag cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggc acc ccc agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttc tca gcc agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gac acc ctg tgc acc agc atc gag ggg cgc atg gat ccc aaa tct tcc 528 Asp Thr Leu Cys Thr Ser Ile Glu Gly Arg Met Asp Pro Lys Ser Ser 165 170 175 gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc ctg ggg 576 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 180 185 190 gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc ctc atg 624 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 195 200 205 atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg agc cac 672 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 210 215 220 gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag gtg 720 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 225 230 235 240 cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc acg tac 768 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 245 250 255 cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aat ggc 816 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 260 265 270 aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc ccc atc 864 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 275 280 285 gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca cag gtg 912 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 290 295 300 tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag gtc agc 960 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 305 310 315 320 ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc gtg gag 1008 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 325 330 335 tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc 1056 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 340 345 350 gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc acc gtg 1104 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 355 360 365 gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc gtg atg 1152 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 370 375 380 cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc ctg tct 1200 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 385 390 395 400 ccg ggt aaa 1209 Pro Gly Lys <210> 102 <211> 403 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of S195-Fc <400> 102 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Ile Glu Gly Arg Met Asp Pro Lys Ser Ser 165 170 175 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 180 185 190 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 195 200 205 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 210 215 220 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 225 230 235 240 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 245 250 255 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 260 265 270 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 275 280 285 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 290 295 300 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 305 310 315 320 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 325 330 335 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 340 345 350 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 355 360 365 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 370 375 380 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 385 390 395 400 Pro Gly Lys <210> 103 <211> 846 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(846) <220> <223> Nucleotide sequence of DcR3 FL-FLAG <400> 103 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac acg cag ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctc tgc ggg gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aac cgt gcc tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ttg gag cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggc acc ccc agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttc tca gcc agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gac acc ctg tgc acc agc tgc act ggc ttc ccc ctc agc acc agg gta 528 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 cca gga gct gag gag tgt gag cgt gcc gtc atc gac ttt gtg gct ttc 576 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 cag gac atc tcc atc aag agg ctg cag cgg ctg ctg cag gcc ctc gag 624 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 gcc ccg gag ggc tgg ggt ccg aca cca agg gcg ggc cgc gcg gcc ttg 672 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 cag ctg aag ctg cgt cgg cgg ctc acg gag ctc ctg ggg gcg cag gac 720 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 ggg gcg ctg ctg gtg cgg ctg ctg cag gcg ctg cgc gtg gcc agg atg 768 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 ccc ggg ctg gag cgg agc gtc cgt gag cgc ttc ctc cct gtg cac ggt 816 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His Gly 260 265 270 acc gca gac tac aag gac gac gat gac aag 846 Thr Ala Asp Tyr Lys Asp Asp Asp Asp Lys 275 280 <210> 104 <211> 282 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of DcR3 FL-FLAG <400> 104 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 Ala Pro Glu Gly Trp Gly Pro Thr Pro Arg Ala Gly Arg Ala Ala Leu 210 215 220 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His Gly 260 265 270 Thr Ala Asp Tyr Lys Asp Asp Asp Asp Lys 275 280 <210> 105 <211> 18 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(18) <220> <223> Nucleotide sequence of Linker IEGRMD <400> 105 atc gag ggg cgc atg gat 18 Ile Glu Gly Arg Met Asp 1 5 <210> 106 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Linker IEGRMD <400> 106 Ile Glu Gly Arg Met Asp 1 5 <210> 107 <211> 498 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(498) <220> <223> Nucleotide sequence of DcR3 from which HBD has been deleted (without a signal peptide) <400> 107 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac acg cag ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctc tgc ggg gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aac cgt gcc tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ttg gag cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggc acc ccc agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttc tca gcc agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gac acc ctg tgc acc agc 498 Asp Thr Leu Cys Thr Ser 165 <210> 108 <211> 166 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of DcR3 from which HBD has been deleted (without a signal peptide) <400> 108 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser 165 <210> 109 <211> 24 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(24) <220> <223> Nucleotide sequence of FLAG tag <400> 109 gac tac aag gac gac gat gac aag 24 Asp Tyr Lys Asp Asp Asp Asp Lys 1 5 <210> 110 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of FLAG tag <400> 110 Asp Tyr Lys Asp Asp Asp Asp Lys 1 5 <210> 111 <211> 813 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(813) <220> <223> Nucleotide sequence of R218Q DcR3 <400> 111 gtg gca gaa aca ccc acc tac ccc tgg cgg gac gca gag aca ggg gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctg gtg tgc gcc cag tgc ccc cca ggc acc ttt gtg cag cgg ccg 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cgc cga gac agc ccc acg acg tgt ggc ccg tgt cca ccg cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac acg cag ttc tgg aac tac cta gag cgc tgc cgc tac tgc aac gtc 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctc tgc ggg gag cgt gag gag gag gca cgg gct tgc cac gcc acc cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aac cgt gcc tgc cgc tgc cgc acc ggc ttc ttc gcg cac gct ggt ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ttg gag cac gca tcg tgt cca cct ggt gcc ggc gtg att gcc ccg 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggc acc ccc agc cag aac acg cag tgc cag ccg tgc ccc cca ggc acc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttc tca gcc agc agc tcc agc tca gag cag tgc cag ccc cac cgc aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgc acg gcc ctg ggc ctg gcc ctc aat gtg cca ggc tct tcc tcc cat 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gac acc ctg tgc acc agc tgc act ggc ttc ccc ctc agc acc agg gta 528 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 cca gga gct gag gag tgt gag cgt gcc gtc atc gac ttt gtg gct ttc 576 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 cag gac atc tcc atc aag agg ctg cag cgg ctg ctg cag gcc ctc gag 624 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 gcc ccg gag ggc tgg ggt ccg aca cca cag gcg ggc cgc gcg gcc ttg 672 Ala Pro Glu Gly Trp Gly Pro Thr Pro Gln Ala Gly Arg Ala Ala Leu 210 215 220 cag ctg aag ctg cgt cgg cgg ctc acg gag ctc ctg ggg gcg cag gac 720 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 ggg gcg ctg ctg gtg cgg ctg ctg cag gcg ctg cgc gtg gcc agg atg 768 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 ccc ggg ctg gag cgg agc gtc cgt gag cgc ttc ctc cct gtg cac 813 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 260 265 270 <210> 112 <211> 271 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of R218Q DcR3 <400> 112 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 Ala Pro Glu Gly Trp Gly Pro Thr Pro Gln Ala Gly Arg Ala Ala Leu 210 215 220 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His 260 265 270 <210> 113 <211> 579 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(579) <220> <223> Nucleotide sequence of soluble recombinant of human LIGHT <400> 113 cat cat cat cat cat cat cat cat cat cat ggt ggt ggt agt ggt ggt 48 His His His His His His His His His His Gly Gly Gly Ser Gly Gly 1 5 10 15 ggt agt ggt ggt ggt agt atc gag gga cgg gac gga cct gca ggc tcc 96 Gly Ser Gly Gly Gly Ser Ile Glu Gly Arg Asp Gly Pro Ala Gly Ser 20 25 30 tgg gag cag ctg ata caa gag cga agg tct cac gag gtc aac cca gca 144 Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser His Glu Val Asn Pro Ala 35 40 45 gcg cat ctc aca ggg gcc aac tcc agc ttg acc ggc agc ggg ggg ccg 192 Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro 50 55 60 ctg tta tgg gag act cag ctg ggc ctg gcc ttc ctg agg ggc ctc agc 240 Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser 65 70 75 80 tac cac gat ggg gcc ctt gtg gtc acc aaa gct ggc tac tac tac atc 288 Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile 85 90 95 tac tcc aag gtg cag ctg ggc ggt gtg ggc tgc ccg ctg ggc ctg gcc 336 Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala 100 105 110 agc acc atc acc cac ggc ctc tac aag cgc aca ccc cgc tac ccc gag 384 Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu 115 120 125 gag ctg gag ctg ttg gtc agc cag cag tca ccc tgc gga cgg gcc acc 432 Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr 130 135 140 agc agc tcc cgg gtc tgg tgg gac agc agc ttc ctg ggt ggt gtg gta 480 Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val 145 150 155 160 cac ctg gag gct ggg gag aag gtg gtc gtc cgt gtg ctg gat gaa cgc 528 His Leu Glu Ala Gly Glu Lys Val Val Val Arg Val Leu Asp Glu Arg 165 170 175 ctg gtt cga ctg cgt gat ggt acc cgg tct tac ttc ggg gct ttc atg 576 Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met 180 185 190 gtg 579 Val <210> 114 <211> 193 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of soluble recombinant of human LIGHT <400> 114 His His His His His His His His His His Gly Gly Gly Ser Gly Gly 1 5 10 15 Gly Ser Gly Gly Gly Ser Ile Glu Gly Arg Asp Gly Pro Ala Gly Ser 20 25 30 Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser His Glu Val Asn Pro Ala 35 40 45 Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro 50 55 60 Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser 65 70 75 80 Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile 85 90 95 Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala 100 105 110 Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu 115 120 125 Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr 130 135 140 Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val 145 150 155 160 His Leu Glu Ala Gly Glu Lys Val Val Val Arg Val Leu Asp Glu Arg 165 170 175 Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met 180 185 190 Val <210> 115 <211> 594 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(594) <220> <223> Nucleotide sequence of soluble recombinant of human TL1A <400> 115 cat cat cat cat cat cat ggt ggt ggt agt ggt ggt ggt agt ggt ggt 48 His His His His His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 ggt agt cta aaa gga cag gag ttt gca cct tca cat cag caa gtt tat 96 Gly Ser Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr 20 25 30 gca cct ctt aga gca gac gga gat aag cca agg gca cac ctg aca gtt 144 Ala Pro Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val 35 40 45 gtg aga caa act ccc aca cag cac ttt aaa aat cag ttc cca gct ctg 192 Val Arg Gln Thr Pro Thr Gln His Phe Lys Asn Gln Phe Pro Ala Leu 50 55 60 cac tgg gaa cat gaa cta ggc ctg gcc ttc acc aag aac cga atg aac 240 His Trp Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn 65 70 75 80 tat acc aac aaa ttc ctg ctg atc cca gag tcg gga gac tac ttc att 288 Tyr Thr Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Ile 85 90 95 tac tcc cag gtc aca ttc cgt ggg atg acc tct gag tgc agt gaa atc 336 Tyr Ser Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile 100 105 110 aga caa gca ggc cga cca aac aag cca gac tcc atc act gtg gtc atc 384 Arg Gln Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile 115 120 125 acc aag gta aca gac agc tac cct gag cca acc cag ctc ctc atg ggg 432 Thr Lys Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly 130 135 140 acc aag tct gta tgc gaa gta ggt agc aac tgg ttc cag ccc atc tac 480 Thr Lys Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr 145 150 155 160 ctc gga gcc atg ttc tcc ttg caa gaa ggg gac aag cta atg gtg aac 528 Leu Gly Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn 165 170 175 gtc agt gac atc tct ttg gtg gat tac aca aaa gaa gat aaa acc ttc 576 Val Ser Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe 180 185 190 ttt gga gcc ttc tta cta 594 Phe Gly Ala Phe Leu Leu 195 <210> 116 <211> 198 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of soluble recombinant of human TL1A <400> 116 His His His His His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 Gly Ser Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr 20 25 30 Ala Pro Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val 35 40 45 Val Arg Gln Thr Pro Thr Gln His Phe Lys Asn Gln Phe Pro Ala Leu 50 55 60 His Trp Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn 65 70 75 80 Tyr Thr Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Ile 85 90 95 Tyr Ser Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile 100 105 110 Arg Gln Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile 115 120 125 Thr Lys Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly 130 135 140 Thr Lys Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr 145 150 155 160 Leu Gly Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn 165 170 175 Val Ser Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe 180 185 190 Phe Gly Ala Phe Leu Leu 195 <210> 117 <211> 462 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(462) <220> <223> Nucleotide sequence of soluble recombinant of human FasL <400> 117 cat cat cat cat cat cat ccc agt cca ccc cct gaa aaa aag gag ctg 48 His His His His His His Pro Ser Pro Pro Glu Lys Lys Glu Leu 1 5 10 15 agg aaa gtg gcc cat tta aca ggc aag tcc aac tca agg tcc atg cct 96 Arg Lys Val Ala His Leu Thr Gly Lys Ser Asn Ser Arg Ser Met Pro 20 25 30 ctg gaa tgg gaa gac acc tat gga att gtc ctg ctt tct gga gtg aag 144 Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu Leu Ser Gly Val Lys 35 40 45 tat aag aag ggt ggc ctt gtg atc aat gaa act ggg ctg tac ttt gta 192 Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly Leu Tyr Phe Val 50 55 60 tat tcc aaa gta tac ttc cgg ggt caa tct tgc aac aac ctg ccc ctg 240 Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Asn Asn Leu Pro Leu 65 70 75 80 agc cac aag gtc tac atg agg aac tct aag tat ccc cag gat ctg gtg 288 Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val 85 90 95 atg atg gag ggg aag atg atg agc tac tgc act act ggg cag atg tgg 336 Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr Gly Gin Met Trp 100 105 110 gcc cgc agc agc tac ctg ggg gca gtg ttc aat ctt acc agt gct gat 384 Ala Arg Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 115 120 125 cat tta tat gtc aac gta tct gag ctc tct ctg gtc aat ttt gag gaa 432 His Leu Tyr Val Asn Val Ser Glu Leu Ser Leu Val Asn Phe Glu Glu 130 135 140 tct cag acg ttt ttc ggc tta tat aag ctc 462 Ser Gln Thr Phe Phe Gly Leu Tyr Lys Leu 145 150 <210> 118 <211> 154 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of soluble recombinant of human FasL <400> 118 His His His His His His Pro Ser Pro Pro Glu Lys Lys Glu Leu 1 5 10 15 Arg Lys Val Ala His Leu Thr Gly Lys Ser Asn Ser Arg Ser Met Pro 20 25 30 Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu Leu Ser Gly Val Lys 35 40 45 Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly Leu Tyr Phe Val 50 55 60 Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Asn Asn Leu Pro Leu 65 70 75 80 Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val 85 90 95 Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr Gly Gin Met Trp 100 105 110 Ala Arg Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 115 120 125 His Leu Tyr Val Asn Val Ser Glu Leu Ser Leu Val Asn Phe Glu Glu 130 135 140 Ser Gln Thr Phe Phe Gly Leu Tyr Lys Leu 145 150 <210> 119 <211> 579 <212> DNA <213> Macaca fascicularis <220> <221> CDS <222> (1)..(579) <220> <223> Nucleotide sequence of soluble recombinant of Macaca fascicularis LIGHT <400> 119 cat cat cat cat cat cat cat cat cat cat ggt ggt ggt agt ggt ggt 48 His His His His His His His His His His Gly Gly Gly Ser Gly Gly 1 5 10 15 ggt agt ggt ggt ggt agt atc gag gga cgg gac gga cac gga ggc tcc 96 Gly Ser Gly Gly Gly Ser Ile Glu Gly Arg Asp Gly His Gly Gly Ser 20 25 30 tgg gag cag ctg ata caa gag cga agg tct cac cag aat aac cca gca 144 Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser His Gln Asn Asn Pro Ala 35 40 45 gca cat ctc aca ggg gcc aac tcc agc ttg acg ggc agc ggg gga ccg 192 Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro 50 55 60 ctg cta tgg gag acc cag ctg ggc ctg gcc ttc ctg agg ggc ctc gac 240 Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Asp 65 70 75 80 tac cac gat ggg gcc ctt gtg gcc aac aag gct ggc tac tac tac atc 288 Tyr His Asp Gly Ala Leu Val Ala Asn Lys Ala Gly Tyr Tyr Tyr Ile 85 90 95 tac tcc aag gtg cag ctg ggc ggt gtg ggc tgc ccg ctg ggc ctg gcc 336 Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala 100 105 110 agc acc atc acc cac ggc ctc tac aag cgc acg ccc cgc tac ccc gag 384 Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu 115 120 125 gag ctg gag ctg ttg gtc agc cag cag tca ccc tgc gga cgg gcc acc 432 Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr 130 135 140 agc aac tcc cgc gtc tgg tgg gac agc agc ttc ctg ggt ggt gtg gtg 480 Ser Asn Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val 145 150 155 160 cac ctg gag gtt ggg gag gag gtg gtc gtc cgt gtg ctg gat gag cgc 528 His Leu Glu Val Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg 165 170 175 ctg gtc cga ctg cgt gat ggc acc cgg tct tac ttc ggg gct ttc atg 576 Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met 180 185 190 gtg 579 Val <210> 120 <211> 193 <212> PRT <213> Macaca fascicularis <220> <223> Amino acid sequence of soluble recombinant of Macaca fascicularis LIGHT <400> 120 His His His His His His His His His His Gly Gly Gly Ser Gly Gly 1 5 10 15 Gly Ser Gly Gly Gly Ser Ile Glu Gly Arg Asp Gly His Gly Gly Ser 20 25 30 Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser His Gln Asn Asn Pro Ala 35 40 45 Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro 50 55 60 Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Asp 65 70 75 80 Tyr His Asp Gly Ala Leu Val Ala Asn Lys Ala Gly Tyr Tyr Tyr Ile 85 90 95 Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala 100 105 110 Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu 115 120 125 Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr 130 135 140 Ser Asn Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val 145 150 155 160 His Leu Glu Val Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg 165 170 175 Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met 180 185 190 Val <210> 121 <211> 594 <212> DNA <213> Macaca fascicularis <220> <221> CDS <222> (1)..(594) <220> <223> Nucleotide sequence of soluble recombinant of Macaca fascicularis TL1A <400> 121 cat cat cat cat cat cat ggt ggt ggt agt ggt ggt ggt agt ggt ggt 48 His His His His His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 ggt agt cta aaa gga cag gag ttt gca cct tca cat cag caa gtt tat 96 Gly Ser Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr 20 25 30 gca cct ctt aga gca gat gga gat aag cca agg gca cac ctg aca gtt 144 Ala Pro Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val 35 40 45 gtg aga caa act ccc aca cag cac tta aaa aat cag ttc cca gct ctg 192 Val Arg Gln Thr Pro Thr Gln His Leu Lys Asn Gln Phe Pro Ala Leu 50 55 60 cac tgg gaa cat gaa cta ggc ctg gcc ttc acc aag aac cga atg aac 240 His Trp Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn 65 70 75 80 tat acc aac aaa ttc ctg ctg atc cca gag tcg gga gac tac ttc gtt 288 Tyr Thr Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Val 85 90 95 tac tcc cag gtc aca ttc cgt ggg atg acc tct gag tgc agt gaa atc 336 Tyr Ser Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile 100 105 110 aga caa gca ggc cga cca aac aag cca gac tcc atc act gtg gtc atc 384 Arg Gln Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile 115 120 125 acc aag gta aca gac agc tac cct gag cca acc cag ctc ctc atg ggg 432 Thr Lys Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly 130 135 140 acc aag tct gtg tgt gaa gta ggc agt aac tgg ttc cag ccc atc tac 480 Thr Lys Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr 145 150 155 160 ctc gga gcc atg ttc tcc ttg caa gaa ggg gac aag ctc atg gtg aac 528 Leu Gly Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn 165 170 175 gtc agt gac atc tct ttg gtg gat tac aca aaa gaa gat aaa acc ttc 576 Val Ser Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe 180 185 190 ttt gga gcc ttc tta cta 594 Phe Gly Ala Phe Leu Leu 195 <210> 122 <211> 198 <212> PRT <213> Macaca fascicularis <220> <223> Amino acid sequence of soluble recombinant of Macaca fascicularis TL1A <400> 122 His His His His His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 Gly Ser Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr 20 25 30 Ala Pro Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val 35 40 45 Val Arg Gln Thr Pro Thr Gln His Leu Lys Asn Gln Phe Pro Ala Leu 50 55 60 His Trp Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn 65 70 75 80 Tyr Thr Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Val 85 90 95 Tyr Ser Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile 100 105 110 Arg Gln Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile 115 120 125 Thr Lys Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly 130 135 140 Thr Lys Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr 145 150 155 160 Leu Gly Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn 165 170 175 Val Ser Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe 180 185 190 Phe Gly Ala Phe Leu Leu 195 <210> 123 <211> 462 <212> DNA <213> Macaca fascicularis <220> <221> CDS <222> (1)..(462) <220> <223> Nucleotide sequence of soluble recombinant of Macaca fascicularis FasL <400> 123 cat cat cat cat cat cat ccc agt cca ccc cct gag aaa aag gag cag 48 His His His His His His Pro Ser Pro Pro Pro Glu Lys Lys Glu Gln 1 5 10 15 agg aaa gtg gcc cat tta aca ggc aag ccc aac tca agg tcc atg cct 96 Arg Lys Val Ala His Leu Thr Gly Lys Pro Asn Ser Arg Ser Met Pro 20 25 30 ctg gaa tgg gaa gac acc tat gga att gtc ctg ctt tct gga gtg aag 144 Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu Leu Ser Gly Val Lys 35 40 45 tat aag aag ggt ggc ctt gtg atc aat gaa act ggg ctg tac ttt gta 192 Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly Leu Tyr Phe Val 50 55 60 tat tcc aaa gta tac ttc cgg ggt caa tct tgc acc aac ctg ccc ctg 240 Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Thr Asn Leu Pro Leu 65 70 75 80 agc cac aag gtc tac atg agg aac tct aag tat ccc cag gat ctg gtg 288 Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val 85 90 95 atg atg gag ggg aag atg atg agc tac tgc act act ggg cag atg tgg 336 Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr Gly Gin Met Trp 100 105 110 gcc cac agc agc tac ctg ggg gca gtg ttc aat ctt acc agt gct gat 384 Ala His Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 115 120 125 cat tta tat gtc aat gta tct gag ctc tct ctg gtc aat ttt gag gaa 432 His Leu Tyr Val Asn Val Ser Glu Leu Ser Leu Val Asn Phe Glu Glu 130 135 140 tct cag aca ttt ttt ggc cta tat aag ctc 462 Ser Gln Thr Phe Phe Gly Leu Tyr Lys Leu 145 150 <210> 124 <211> 154 <212> PRT <213> Macaca fascicularis <220> <223> Amino acid sequence of soluble recombinant of Macaca fascicularis FasL <400> 124 His His His His His His Pro Ser Pro Pro Pro Glu Lys Lys Glu Gln 1 5 10 15 Arg Lys Val Ala His Leu Thr Gly Lys Pro Asn Ser Arg Ser Met Pro 20 25 30 Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu Leu Ser Gly Val Lys 35 40 45 Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly Leu Tyr Phe Val 50 55 60 Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Thr Asn Leu Pro Leu 65 70 75 80 Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val 85 90 95 Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr Gly Gin Met Trp 100 105 110 Ala His Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 115 120 125 His Leu Tyr Val Asn Val Ser Glu Leu Ser Leu Val Asn Phe Glu Glu 130 135 140 Ser Gln Thr Phe Phe Gly Leu Tyr Lys Leu 145 150 <210> 125 <211> 582 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1)..(582) <220> <223> Nucleotide sequence of soluble recombinant of mouse LIGHT <400> 125 cat cat cat cat cat cat cat cat cat cat ggt ggt ggt agt ggt ggt 48 His His His His His His His His His His Gly Gly Gly Ser Gly Gly 1 5 10 15 ggt agt ggt ggt ggt agt atc gag gga cgg gat gga ggc aaa ggc tcc 96 Gly Ser Gly Gly Gly Ser Ile Glu Gly Arg Asp Gly Gly Lys Gly Ser 20 25 30 tgg gag aag ctg ata caa gat caa cga tct cac cag gcc aac cca gca 144 Trp Glu Lys Leu Ile Gln Asp Gln Arg Ser His Gln Ala Asn Pro Ala 35 40 45 gca cat ctt aca gga gcc aac gcc agc ttg ata ggt att ggt gga cct 192 Ala His Leu Thr Gly Ala Asn Ala Ser Leu Ile Gly Ile Gly Gly Pro 50 55 60 ctg tta tgg gag aca cga ctt ggc ctg gcc ttc ttg agg ggc ttg acg 240 Leu Leu Trp Glu Thr Arg Leu Gly Leu Ala Phe Leu Arg Gly Leu Thr 65 70 75 80 tat cat gat ggg gcc ctg gtg acc atg gag ccc ggt tac tac tat gtg 288 Tyr His Asp Gly Ala Leu Val Thr Met Glu Pro Gly Tyr Tyr Tyr Val 85 90 95 tac tcc aaa gtg cag ctg agc ggc gtg ggc tgc ccc cag ggg ctg gcc 336 Tyr Ser Lys Val Gln Leu Ser Gly Val Gly Cys Pro Gln Gly Leu Ala 100 105 110 aat ggc ctc ccc atc acc cat gga cta tac aag cgc aca tcc cgc tac 384 Asn Gly Leu Pro Ile Thr His Gly Leu Tyr Lys Arg Thr Ser Arg Tyr 115 120 125 ccg aag gag tta gaa ctg ctg gtc agt cgg cgg tca ccc tgt ggc cgg 432 Pro Lys Glu Leu Glu Leu Leu Val Ser Arg Arg Ser Pro Cys Gly Arg 130 135 140 gcc aac agc tcc cga gtc tgg tgg gac agc agc ttc ctg ggc ggc gtg 480 Ala Asn Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val 145 150 155 160 gta cat ctg gag gct ggg gaa gag gtg gtg gtc cgc gtg cct gga aac 528 Val His Leu Glu Ala Gly Glu Glu Val Val Val Val Arg Val Pro Gly Asn 165 170 175 cgc ctg gtc aga cca cgt gac ggc acc agg tcc tat ttc gga gct ttc 576 Arg Leu Val Arg Pro Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe 180 185 190 atg gtc 582 Met Val <210> 126 <211> 194 <212> PRT <213> Mus musculus <220> <223> Amino acid sequence of soluble recombinant of mouse LIGHT <400> 126 His His His His His His His His His His Gly Gly Gly Ser Gly Gly 1 5 10 15 Gly Ser Gly Gly Gly Ser Ile Glu Gly Arg Asp Gly Gly Lys Gly Ser 20 25 30 Trp Glu Lys Leu Ile Gln Asp Gln Arg Ser His Gln Ala Asn Pro Ala 35 40 45 Ala His Leu Thr Gly Ala Asn Ala Ser Leu Ile Gly Ile Gly Gly Pro 50 55 60 Leu Leu Trp Glu Thr Arg Leu Gly Leu Ala Phe Leu Arg Gly Leu Thr 65 70 75 80 Tyr His Asp Gly Ala Leu Val Thr Met Glu Pro Gly Tyr Tyr Tyr Val 85 90 95 Tyr Ser Lys Val Gln Leu Ser Gly Val Gly Cys Pro Gln Gly Leu Ala 100 105 110 Asn Gly Leu Pro Ile Thr His Gly Leu Tyr Lys Arg Thr Ser Arg Tyr 115 120 125 Pro Lys Glu Leu Glu Leu Leu Val Ser Arg Arg Ser Pro Cys Gly Arg 130 135 140 Ala Asn Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val 145 150 155 160 Val His Leu Glu Ala Gly Glu Glu Val Val Val Val Arg Val Pro Gly Asn 165 170 175 Arg Leu Val Arg Pro Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe 180 185 190 Met Val <210> 127 <211> 585 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1)..(585) <220> <223> Nucleotide sequence of soluble recombinant of mouse TL1A <400> 127 cat cat cat cat cat cat ggt ggt ggt agt ggt ggt ggt agt ggt ggt 48 His His His His His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 ggt agt ata aca gaa gag agg tct gag cct tca cca cag caa gtt tac 96 Gly Ser Ile Thr Glu Glu Arg Ser Glu Pro Ser Pro Gln Gln Val Tyr 20 25 30 tca cct ccc aga ggc aag ccg aga gca cac ctg aca att aag aaa caa 144 Ser Pro Pro Arg Gly Lys Pro Arg Ala His Leu Thr Ile Lys Lys Gln 35 40 45 acc cca gca cca cat ctg aaa aat cag ctc tct gct cta cac tgg gaa 192 Thr Pro Ala Pro His Leu Lys Asn Gln Leu Ser Ala Leu His Trp Glu 50 55 60 cat gac cta ggg atg gcc ttc acc aag aac ggg atg aag tac atc aac 240 His Asp Leu Gly Met Ala Phe Thr Lys Asn Gly Met Lys Tyr Ile Asn 65 70 75 80 aaa tcc ctg gtg atc cca gag tca gga gac tat ttc atc tac tcc cag 288 Lys Ser Leu Val Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser Gln 85 90 95 atc aca ttc cga ggg acc aca tct gtg tgt ggt gac atc agt cgg ggg 336 Ile Thr Phe Arg Gly Thr Thr Ser Val Cys Gly Asp Ile Ser Arg Gly 100 105 110 aga cga cca aac aag cca gac tcc atc acc atg gtt atc acc aag gta 384 Arg Arg Pro Asn Lys Pro Asp Ser Ile Thr Met Val Ile Thr Lys Val 115 120 125 gca gac agc tac cct gag cct gcc cgc cta cta aca ggg tcc aag tct 432 Ala Asp Ser Tyr Pro Glu Pro Ala Arg Leu Leu Thr Gly Ser Lys Ser 130 135 140 gtg tgt gaa ata agc aac aac tgg ttc cag tcc ctc tac ctt ggg gcc 480 Val Cys Glu Ile Ser Asn Asn Trp Phe Gln Ser Leu Tyr Leu Gly Ala 145 150 155 160 acg ttc tcc ttg gaa gaa gga gac aga cta atg gta aac gtc agt gac 528 Thr Phe Ser Leu Glu Glu Gly Asp Arg Leu Met Val Asn Val Ser Asp 165 170 175 atc tcc ttg gtg gat tac aca aaa gaa gat aaa act ttc ttt gga gct 576 Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly Ala 180 185 190 ttc ttg cta 585 Phe Leu Leu 195 <210> 128 <211> 195 <212> PRT <213> Mus musculus <220> <223> Amino acid sequence of soluble recombinant of mouse TL1A <400> 128 His His His His His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 Gly Ser Ile Thr Glu Glu Arg Ser Glu Pro Ser Pro Gln Gln Val Tyr 20 25 30 Ser Pro Pro Arg Gly Lys Pro Arg Ala His Leu Thr Ile Lys Lys Gln 35 40 45 Thr Pro Ala Pro His Leu Lys Asn Gln Leu Ser Ala Leu His Trp Glu 50 55 60 His Asp Leu Gly Met Ala Phe Thr Lys Asn Gly Met Lys Tyr Ile Asn 65 70 75 80 Lys Ser Leu Val Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser Gln 85 90 95 Ile Thr Phe Arg Gly Thr Thr Ser Val Cys Gly Asp Ile Ser Arg Gly 100 105 110 Arg Arg Pro Asn Lys Pro Asp Ser Ile Thr Met Val Ile Thr Lys Val 115 120 125 Ala Asp Ser Tyr Pro Glu Pro Ala Arg Leu Leu Thr Gly Ser Lys Ser 130 135 140 Val Cys Glu Ile Ser Asn Asn Trp Phe Gln Ser Leu Tyr Leu Gly Ala 145 150 155 160 Thr Phe Ser Leu Glu Glu Gly Asp Arg Leu Met Val Asn Val Ser Asp 165 170 175 Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly Ala 180 185 190 Phe Leu Leu 195 <210> 129 <211> 462 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1)..(462) <220> <223> Nucleotide sequence of soluble recombinant of mouse FasL <400> 129 cat cat cat cat cat cat ccc agt aca ccc tct gaa aaa aaa gag ccg 48 His His His His His His Pro Ser Thr Pro Ser Glu Lys Lys Glu Pro 1 5 10 15 agg agt gtg gcc cat tta aca ggg aac ccc cac tca agg tcc atc cct 96 Arg Ser Val Ala His Leu Thr Gly Asn Pro His Ser Arg Ser Ile Pro 20 25 30 ctg gaa tgg gaa gac aca tat gga acc gct ctg atc tct gga gtg aag 144 Leu Glu Trp Glu Asp Thr Tyr Gly Thr Ala Leu Ile Ser Gly Val Lys 35 40 45 tat aag aaa ggt ggc ctt gtg atc aac gaa act ggg ttg tac ttc gtg 192 Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly Leu Tyr Phe Val 50 55 60 tat tcc aaa gta tac ttc cgg ggt cag tct tgc aac aac cag ccc cta 240 Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Asn Asn Gln Pro Leu 65 70 75 80 aac cac aag gtc tat atg agg aac tct aag tat cct gag gat ctg gtg 288 Asn His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Glu Asp Leu Val 85 90 95 cta atg gag gag aag agg ttg aac tac tgc act act gga cag ata tgg 336 Leu Met Glu Glu Lys Arg Leu Asn Tyr Cys Thr Thr Gly Gln Ile Trp 100 105 110 gcc cac agc agc tac ctg ggg gca gta ttc aat ctt acc agt gct gac 384 Ala His Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 115 120 125 cat tta tat gtc aac ata tct caa ctc tct ctg atc aat ttt gag gaa 432 His Leu Tyr Val Asn Ile Ser Gln Leu Ser Leu Ile Asn Phe Glu Glu 130 135 140 tct aag acc ttt ttc ggc tta tat aag ctc 462 Ser Lys Thr Phe Phe Gly Leu Tyr Lys Leu 145 150 <210> 130 <211> 154 <212> PRT <213> Mus musculus <220> <223> Amino acid sequence of soluble recombinant of mouse FasL <400> 130 His His His His His His Pro Ser Thr Pro Ser Glu Lys Lys Glu Pro 1 5 10 15 Arg Ser Val Ala His Leu Thr Gly Asn Pro His Ser Arg Ser Ile Pro 20 25 30 Leu Glu Trp Glu Asp Thr Tyr Gly Thr Ala Leu Ile Ser Gly Val Lys 35 40 45 Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly Leu Tyr Phe Val 50 55 60 Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Asn Asn Gln Pro Leu 65 70 75 80 Asn His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro Glu Asp Leu Val 85 90 95 Leu Met Glu Glu Lys Arg Leu Asn Tyr Cys Thr Thr Gly Gln Ile Trp 100 105 110 Ala His Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu Thr Ser Ala Asp 115 120 125 His Leu Tyr Val Asn Ile Ser Gln Leu Ser Leu Ile Asn Phe Glu Glu 130 135 140 Ser Lys Thr Phe Phe Gly Leu Tyr Lys Leu 145 150 <210> 131 <211> 501 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(501) <220> <223> Nucleotide sequence of extracellular region of human LIGHT (Asp74-Val240) <400> 131 gac gga cct gca ggc tcc tgg gag cag ctg ata caa gag cga agg tct 48 Asp Gly Pro Ala Gly Ser Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser 1 5 10 15 cac gag gtc aac cca gca gcg cat ctc aca ggg gcc aac tcc agc ttg 96 His Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu 20 25 30 acc ggc agc ggg ggg ccg ctg tta tgg gag act cag ctg ggc ctg gcc 144 Thr Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala 35 40 45 ttc ctg agg ggc ctc agc tac cac gat ggg gcc ctt gtg gtc acc aaa 192 Phe Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys 50 55 60 gct ggc tac tac tac atc tac tcc aag gtg cag ctg ggc ggt gtg ggc 240 Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly 65 70 75 80 tgc ccg ctg ggc ctg gcc agc acc atc acc cac ggc ctc tac aag cgc 288 Cys Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg 85 90 95 aca ccc cgc tac ccc gag gag ctg gag ctg ttg gtc agc cag cag tca 336 Thr Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser 100 105 110 ccc tgc gga cgg gcc acc agc agc tcc cgg gtc tgg tgg gac agc agc 384 Pro Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser 115 120 125 ttc ctg ggt ggt gtg gta cac ctg gag gct ggg gag aag gtg gtc gtc 432 Phe Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Lys Val Val Val 130 135 140 cgt gtg ctg gat gaa cgc ctg gtt cga ctg cgt gat ggt acc cgg tct 480 Arg Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser 145 150 155 160 tac ttc ggg gct ttc atg gtg 501 Tyr Phe Gly Ala Phe Met Val 165 <210> 132 <211> 167 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of extracellular region of human LIGHT (Asp74-Val240) <400> 132 Asp Gly Pro Ala Gly Ser Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser 1 5 10 15 His Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu 20 25 30 Thr Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala 35 40 45 Phe Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys 50 55 60 Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly 65 70 75 80 Cys Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg 85 90 95 Thr Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser 100 105 110 Pro Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser 115 120 125 Phe Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Lys Val Val Val 130 135 140 Arg Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser 145 150 155 160 Tyr Phe Gly Ala Phe Met Val 165 <210> 133 <211> 501 <212> DNA <213> Macaca fascicularis <220> <221> CDS <222> (1)..(501) <220> <223> Nucleotide sequence of extracellular region of Macaca fascicularis LIGHT (Asp74-Val240) <400> 133 gac gga cac gga ggc tcc tgg gag cag ctg ata caa gag cga agg tct 48 Asp Gly His Gly Gly Ser Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser 1 5 10 15 cac cag aat aac cca gca gca cat ctc aca ggg gcc aac tcc agc ttg 96 His Gln Asn Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu 20 25 30 acg ggc agc ggg gga ccg ctg cta tgg gag acc cag ctg ggc ctg gcc 144 Thr Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala 35 40 45 ttc ctg agg ggc ctc gac tac cac gat ggg gcc ctt gtg gcc aac aag 192 Phe Leu Arg Gly Leu Asp Tyr His Asp Gly Ala Leu Val Ala Asn Lys 50 55 60 gct ggc tac tac tac atc tac tcc aag gtg cag ctg ggc ggt gtg ggc 240 Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly 65 70 75 80 tgc ccg ctg ggc ctg gcc agc acc atc acc cac ggc ctc tac aag cgc 288 Cys Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg 85 90 95 acg ccc cgc tac ccc gag gag ctg gag ctg ttg gtc agc cag cag tca 336 Thr Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser 100 105 110 ccc tgc gga cgg gcc acc agc aac tcc cgc gtc tgg tgg gac agc agc 384 Pro Cys Gly Arg Ala Thr Ser Asn Ser Arg Val Trp Trp Asp Ser Ser 115 120 125 ttc ctg ggt ggt gtg gtg cac ctg gag gtt ggg gag gag gtg gtc gtc 432 Phe Leu Gly Gly Val Val His Leu Glu Val Gly Glu Glu Val Val Val 130 135 140 cgt gtg ctg gat gag cgc ctg gtc cga ctg cgt gat ggc acc cgg tct 480 Arg Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser 145 150 155 160 tac ttc ggg gct ttc atg gtg 501 Tyr Phe Gly Ala Phe Met Val 165 <210> 134 <211> 167 <212> PRT <213> Macaca fascicularis <220> <223> Amino acid sequence of extracellular region of Macaca fascicularis LIGHT (Asp74-Val240) <400> 134 Asp Gly His Gly Gly Ser Trp Glu Gln Leu Ile Gln Glu Arg Arg Ser 1 5 10 15 His Gln Asn Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu 20 25 30 Thr Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala 35 40 45 Phe Leu Arg Gly Leu Asp Tyr His Asp Gly Ala Leu Val Ala Asn Lys 50 55 60 Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly 65 70 75 80 Cys Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg 85 90 95 Thr Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser 100 105 110 Pro Cys Gly Arg Ala Thr Ser Asn Ser Arg Val Trp Trp Asp Ser Ser 115 120 125 Phe Leu Gly Gly Val Val His Leu Glu Val Gly Glu Glu Val Val Val 130 135 140 Arg Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser 145 150 155 160 Tyr Phe Gly Ala Phe Met Val 165 <210> 135 <211> 504 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1)..(504) <220> <223> Nucleotide sequence of extracellular region of mouse LIGHT (Asp74-Val240) <400> 135 gat gga ggc aaa ggc tcc tgg gag aag ctg ata caa gat caa cga tct 48 Asp Gly Gly Lys Gly Ser Trp Glu Lys Leu Ile Gln Asp Gln Arg Ser 1 5 10 15 cac cag gcc aac cca gca gca cat ctt aca gga gcc aac gcc agc ttg 96 His Gln Ala Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ala Ser Leu 20 25 30 ata ggt att ggt gga cct ctg tta tgg gag aca cga ctt ggc ctg gcc 144 Ile Gly Ile Gly Gly Pro Leu Leu Trp Glu Thr Arg Leu Gly Leu Ala 35 40 45 ttc ttg agg ggc ttg acg tat cat gat ggg gcc ctg gtg acc atg gag 192 Phe Leu Arg Gly Leu Thr Tyr His Asp Gly Ala Leu Val Thr Met Glu 50 55 60 ccc ggt tac tac tat gtg tac tcc aaa gtg cag ctg agc ggc gtg ggc 240 Pro Gly Tyr Tyr Tyr Val Tyr Ser Lys Val Gln Leu Ser Gly Val Gly 65 70 75 80 tgc ccc cag ggg ctg gcc aat ggc ctc ccc atc acc cat gga cta tac 288 Cys Pro Gln Gly Leu Ala Asn Gly Leu Pro Ile Thr His Gly Leu Tyr 85 90 95 aag cgc aca tcc cgc tac ccg aag gag tta gaa ctg ctg gtc agt cgg 336 Lys Arg Thr Ser Arg Tyr Pro Lys Glu Leu Glu Leu Leu Val Ser Arg 100 105 110 cgg tca ccc tgt ggc cgg gcc aac agc tcc cga gtc tgg tgg gac agc 384 Arg Ser Pro Cys Gly Arg Ala Asn Ser Ser Arg Val Trp Trp Asp Ser 115 120 125 agc ttc ctg ggc ggc gtg gta cat ctg gag gct ggg gaa gag gtg gtg 432 Ser Phe Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val 130 135 140 gtc cgc gtg cct gga aac cgc ctg gtc aga cca cgt gac ggc acc agg 480 Val Arg Val Pro Gly Asn Arg Leu Val Arg Pro Arg Asp Gly Thr Arg 145 150 155 160 tcc tat ttc gga gct ttc atg gtc 504 Ser Tyr Phe Gly Ala Phe Met Val 165 <210> 136 <211> 168 <212> PRT <213> Mus musculus <220> <223> Amino acid sequence of extracellular region of mouse LIGHT (Asp74-Val240) <400> 136 Asp Gly Gly Lys Gly Ser Trp Glu Lys Leu Ile Gln Asp Gln Arg Ser 1 5 10 15 His Gln Ala Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ala Ser Leu 20 25 30 Ile Gly Ile Gly Gly Pro Leu Leu Trp Glu Thr Arg Leu Gly Leu Ala 35 40 45 Phe Leu Arg Gly Leu Thr Tyr His Asp Gly Ala Leu Val Thr Met Glu 50 55 60 Pro Gly Tyr Tyr Tyr Val Tyr Ser Lys Val Gln Leu Ser Gly Val Gly 65 70 75 80 Cys Pro Gln Gly Leu Ala Asn Gly Leu Pro Ile Thr His Gly Leu Tyr 85 90 95 Lys Arg Thr Ser Arg Tyr Pro Lys Glu Leu Glu Leu Leu Val Ser Arg 100 105 110 Arg Ser Pro Cys Gly Arg Ala Asn Ser Ser Arg Val Trp Trp Asp Ser 115 120 125 Ser Phe Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val 130 135 140 Val Arg Val Pro Gly Asn Arg Leu Val Arg Pro Arg Asp Gly Thr Arg 145 150 155 160 Ser Tyr Phe Gly Ala Phe Met Val 165 <210> 137 <211> 540 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(540) <220> <223> Nucleotide sequence of extracellular region of human TL1A (Leu72-Leu251) <400> 137 cta aaa gga cag gag ttt gca cct tca cat cag caa gtt tat gca cct 48 Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr Ala Pro 1 5 10 15 ctt aga gca gac gga gat aag cca agg gca cac ctg aca gtt gtg aga 96 Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val Val Arg 20 25 30 caa act ccc aca cag cac ttt aaa aat cag ttc cca gct ctg cac tgg 144 Gln Thr Pro Thr Gln His Phe Lys Asn Gln Phe Pro Ala Leu His Trp 35 40 45 gaa cat gaa cta ggc ctg gcc ttc acc aag aac cga atg aac tat acc 192 Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn Tyr Thr 50 55 60 aac aaa ttc ctg ctg atc cca gag tcg gga gac tac ttc att tac tcc 240 Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser 65 70 75 80 cag gtc aca ttc cgt ggg atg acc tct gag tgc agt gaa atc aga caa 288 Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile Arg Gln 85 90 95 gca ggc cga cca aac aag cca gac tcc atc act gtg gtc atc acc aag 336 Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile Thr Lys 100 105 110 gta aca gac agc tac cct gag cca acc cag ctc ctc atg ggg acc aag 384 Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly Thr Lys 115 120 125 tct gta tgc gaa gta ggt agc aac tgg ttc cag ccc atc tac ctc gga 432 Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr Leu Gly 130 135 140 gcc atg ttc tcc ttg caa gaa ggg gac aag cta atg gtg aac gtc agt 480 Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn Val Ser 145 150 155 160 gac atc tct ttg gtg gat tac aca aaa gaa gat aaa acc ttc ttt gga 528 Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly 165 170 175 gcc ttc tta cta 540 Ala Phe Leu Leu 180 <210> 138 <211> 180 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of extracellular region of human TL1A (Leu72-Leu251) <400> 138 Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr Ala Pro 1 5 10 15 Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val Val Arg 20 25 30 Gln Thr Pro Thr Gln His Phe Lys Asn Gln Phe Pro Ala Leu His Trp 35 40 45 Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn Tyr Thr 50 55 60 Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser 65 70 75 80 Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile Arg Gln 85 90 95 Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile Thr Lys 100 105 110 Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly Thr Lys 115 120 125 Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr Leu Gly 130 135 140 Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn Val Ser 145 150 155 160 Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly 165 170 175 Ala Phe Leu Leu 180 <210> 139 <211> 540 <212> DNA <213> Macaca fascicularis <220> <221> CDS <222> (1)..(540) <220> <223> Nucleotide sequence of extracellular region of Macaca fascicularis TL1A (Leu72-Leu251) <400> 139 cta aaa gga cag gag ttt gca cct tca cat cag caa gtt tat gca cct 48 Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr Ala Pro 1 5 10 15 ctt aga gca gat gga gat aag cca agg gca cac ctg aca gtt gtg aga 96 Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val Val Arg 20 25 30 caa act ccc aca cag cac tta aaa aat cag ttc cca gct ctg cac tgg 144 Gln Thr Pro Thr Gln His Leu Lys Asn Gln Phe Pro Ala Leu His Trp 35 40 45 gaa cat gaa cta ggc ctg gcc ttc acc aag aac cga atg aac tat acc 192 Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn Tyr Thr 50 55 60 aac aaa ttc ctg ctg atc cca gag tcg gga gac tac ttc gtt tac tcc 240 Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Val Tyr Ser 65 70 75 80 cag gtc aca ttc cgt ggg atg acc tct gag tgc agt gaa atc aga caa 288 Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile Arg Gln 85 90 95 gca ggc cga cca aac aag cca gac tcc atc act gtg gtc atc acc aag 336 Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile Thr Lys 100 105 110 gta aca gac agc tac cct gag cca acc cag ctc ctc atg ggg acc aag 384 Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly Thr Lys 115 120 125 tct gtg tgt gaa gta ggc agt aac tgg ttc cag ccc atc tac ctc gga 432 Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr Leu Gly 130 135 140 gcc atg ttc tcc ttg caa gaa ggg gac aag ctc atg gtg aac gtc agt 480 Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn Val Ser 145 150 155 160 gac atc tct ttg gtg gat tac aca aaa gaa gat aaa acc ttc ttt gga 528 Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly 165 170 175 gcc ttc tta cta 540 Ala Phe Leu Leu 180 <210> 140 <211> 180 <212> PRT <213> Macaca fascicularis <220> <223> Amino acid sequence of extracellular region of Macaca fascicularis TL1A (Leu72-Leu251) <400> 140 Leu Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr Ala Pro 1 5 10 15 Leu Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val Val Arg 20 25 30 Gln Thr Pro Thr Gln His Leu Lys Asn Gln Phe Pro Ala Leu His Trp 35 40 45 Glu His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn Tyr Thr 50 55 60 Asn Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Val Tyr Ser 65 70 75 80 Gln Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile Arg Gln 85 90 95 Ala Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile Thr Lys 100 105 110 Val Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly Thr Lys 115 120 125 Ser Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr Leu Gly 130 135 140 Ala Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn Val Ser 145 150 155 160 Asp Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly 165 170 175 Ala Phe Leu Leu 180 <210> 141 <211> 531 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1)..(531) <220> <223> Nucleotide sequence of extracellular region of mouse TL1A (Ile94-Leu270) <400> 141 ata aca gaa gag agg tct gag cct tca cca cag caa gtt tac tca cct 48 Ile Thr Glu Glu Arg Ser Glu Pro Ser Pro Gln Gln Val Tyr Ser Pro 1 5 10 15 ccc aga ggc aag ccg aga gca cac ctg aca att aag aaa caa acc cca 96 Pro Arg Gly Lys Pro Arg Ala His Leu Thr Ile Lys Lys Gln Thr Pro 20 25 30 gca cca cat ctg aaa aat cag ctc tct gct cta cac tgg gaa cat gac 144 Ala Pro His Leu Lys Asn Gln Leu Ser Ala Leu His Trp Glu His Asp 35 40 45 cta ggg atg gcc ttc acc aag aac ggg atg aag tac atc aac aaa tcc 192 Leu Gly Met Ala Phe Thr Lys Asn Gly Met Lys Tyr Ile Asn Lys Ser 50 55 60 ctg gtg atc cca gag tca gga gac tat ttc atc tac tcc cag atc aca 240 Leu Val Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser Gln Ile Thr 65 70 75 80 ttc cga ggg acc aca tct gtg tgt ggt gac atc agt cgg ggg aga cga 288 Phe Arg Gly Thr Thr Ser Val Cys Gly Asp Ile Ser Arg Gly Arg Arg 85 90 95 cca aac aag cca gac tcc atc acc atg gtt atc acc aag gta gca gac 336 Pro Asn Lys Pro Asp Ser Ile Thr Met Val Ile Thr Lys Val Ala Asp 100 105 110 agc tac cct gag cct gcc cgc cta cta aca ggg tcc aag tct gtg tgt 384 Ser Tyr Pro Glu Pro Ala Arg Leu Leu Thr Gly Ser Lys Ser Val Cys 115 120 125 gaa ata agc aac aac tgg ttc cag tcc ctc tac ctt ggg gcc acg ttc 432 Glu Ile Ser Asn Asn Trp Phe Gln Ser Leu Tyr Leu Gly Ala Thr Phe 130 135 140 tcc ttg gaa gaa gga gac aga cta atg gta aac gtc agt gac atc tcc 480 Ser Leu Glu Glu Gly Asp Arg Leu Met Val Asn Val Ser Asp Ile Ser 145 150 155 160 ttg gtg gat tac aca aaa gaa gat aaa act ttc ttt gga gct ttc ttg 528 Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly Ala Phe Leu 165 170 175 cta 531 Leu <210> 142 <211> 177 <212> PRT <213> Mus musculus <220> <223> Amino acid sequence of extracellular region of mouse TL1A (Ile94-Leu270) <400> 142 Ile Thr Glu Glu Arg Ser Glu Pro Ser Pro Gln Gln Val Tyr Ser Pro 1 5 10 15 Pro Arg Gly Lys Pro Arg Ala His Leu Thr Ile Lys Lys Gln Thr Pro 20 25 30 Ala Pro His Leu Lys Asn Gln Leu Ser Ala Leu His Trp Glu His Asp 35 40 45 Leu Gly Met Ala Phe Thr Lys Asn Gly Met Lys Tyr Ile Asn Lys Ser 50 55 60 Leu Val Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser Gln Ile Thr 65 70 75 80 Phe Arg Gly Thr Thr Ser Val Cys Gly Asp Ile Ser Arg Gly Arg Arg 85 90 95 Pro Asn Lys Pro Asp Ser Ile Thr Met Val Ile Thr Lys Val Ala Asp 100 105 110 Ser Tyr Pro Glu Pro Ala Arg Leu Leu Thr Gly Ser Lys Ser Val Cys 115 120 125 Glu Ile Ser Asn Asn Trp Phe Gln Ser Leu Tyr Leu Gly Ala Thr Phe 130 135 140 Ser Leu Glu Glu Gly Asp Arg Leu Met Val Asn Val Ser Asp Ile Ser 145 150 155 160 Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly Ala Phe Leu 165 170 175 Leu <210> 143 <211> 444 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(444) <220> <223> Nucleotide sequence of extracellular region of human FasL (Pro134-Leu281) <400> 143 ccc agt cca ccc cct gaa aaa aag gag ctg agg aaa gtg gcc cat tta 48 Pro Ser Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His Leu 1 5 10 15 aca ggc aag tcc aac tca agg tcc atg cct ctg gaa tgg gaa gac acc 96 Thr Gly Lys Ser Asn Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr 20 25 30 tat gga att gtc ctg ctt tct gga gtg aag tat aag aag ggt ggc ctt 144 Tyr Gly Ile Val Leu Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 35 40 45 gtg atc aat gaa act ggg ctg tac ttt gta tat tcc aaa gta tac ttc 192 Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe 50 55 60 cgg ggt caa tct tgc aac aac ctg ccc ctg agc cac aag gtc tac atg 240 Arg Gly Gln Ser Cys Asn Asn Leu Pro Leu Ser His Lys Val Tyr Met 65 70 75 80 agg aac tct aag tat ccc cag gat ctg gtg atg atg gag ggg aag atg 288 Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val Met Met Glu Gly Lys Met 85 90 95 atg agc tac tgc act act ggg cag atg tgg gcc cgc agc agc tac ctg 336 Met Ser Tyr Cys Thr Thr Gly Gln Met Trp Ala Arg Ser Ser Tyr Leu 100 105 110 ggg gca gtg ttc aat ctt acc agt gct gat cat tta tat gtc aac gta 384 Gly Ala Val Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Val 115 120 125 tct gag ctc tct ctg gtc aat ttt gag gaa tct cag acg ttt ttc ggc 432 Ser Glu Leu Ser Leu Val Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly 130 135 140 tta tat aag ctc 444 Leu Tyr Lys Leu 145 <210> 144 <211> 148 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of extracellular region of human FasL (Pro134-Leu281) <400> 144 Pro Ser Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His Leu 1 5 10 15 Thr Gly Lys Ser Asn Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr 20 25 30 Tyr Gly Ile Val Leu Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 35 40 45 Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe 50 55 60 Arg Gly Gln Ser Cys Asn Asn Leu Pro Leu Ser His Lys Val Tyr Met 65 70 75 80 Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val Met Met Glu Gly Lys Met 85 90 95 Met Ser Tyr Cys Thr Thr Gly Gln Met Trp Ala Arg Ser Ser Tyr Leu 100 105 110 Gly Ala Val Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Val 115 120 125 Ser Glu Leu Ser Leu Val Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly 130 135 140 Leu Tyr Lys Leu 145 <210> 145 <211> 444 <212> DNA <213> Macaca fascicularis <220> <221> CDS <222> (1)..(444) <220> <223> Nucleotide sequence of extracellular region of Macaca fascicularis FasL (Pro133-Leu280) <400> 145 ccc agt cca ccc cct gag aaa aag gag cag agg aaa gtg gcc cat tta 48 Pro Ser Pro Pro Glu Lys Lys Glu Gln Arg Lys Val Ala His Leu 1 5 10 15 aca ggc aag ccc aac tca agg tcc atg cct ctg gaa tgg gaa gac acc 96 Thr Gly Lys Pro Asn Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr 20 25 30 tat gga att gtc ctg ctt tct gga gtg aag tat aag aag ggt ggc ctt 144 Tyr Gly Ile Val Leu Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 35 40 45 gtg atc aat gaa act ggg ctg tac ttt gta tat tcc aaa gta tac ttc 192 Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe 50 55 60 cgg ggt caa tct tgc acc aac ctg ccc ctg agc cac aag gtc tac atg 240 Arg Gly Gln Ser Cys Thr Asn Leu Pro Leu Ser His Lys Val Tyr Met 65 70 75 80 agg aac tct aag tat ccc cag gat ctg gtg atg atg gag ggg aag atg 288 Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val Met Met Glu Gly Lys Met 85 90 95 atg agc tac tgc act act ggg cag atg tgg gcc cac agc agc tac ctg 336 Met Ser Tyr Cys Thr Thr Gly Gln Met Trp Ala His Ser Ser Tyr Leu 100 105 110 ggg gca gtg ttc aat ctt acc agt gct gat cat tta tat gtc aat gta 384 Gly Ala Val Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Val 115 120 125 tct gag ctc tct ctg gtc aat ttt gag gaa tct cag aca ttt ttt ggc 432 Ser Glu Leu Ser Leu Val Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly 130 135 140 cta tat aag ctc 444 Leu Tyr Lys Leu 145 <210> 146 <211> 148 <212> PRT <213> Macaca fascicularis <220> <223> Nucleotide sequence of extracellular region of Macaca fascicularis FasL (Pro133-Leu280) <400> 146 Pro Ser Pro Pro Glu Lys Lys Glu Gln Arg Lys Val Ala His Leu 1 5 10 15 Thr Gly Lys Pro Asn Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr 20 25 30 Tyr Gly Ile Val Leu Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 35 40 45 Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe 50 55 60 Arg Gly Gln Ser Cys Thr Asn Leu Pro Leu Ser His Lys Val Tyr Met 65 70 75 80 Arg Asn Ser Lys Tyr Pro Gln Asp Leu Val Met Met Glu Gly Lys Met 85 90 95 Met Ser Tyr Cys Thr Thr Gly Gln Met Trp Ala His Ser Ser Tyr Leu 100 105 110 Gly Ala Val Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Val 115 120 125 Ser Glu Leu Ser Leu Val Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly 130 135 140 Leu Tyr Lys Leu 145 <210> 147 <211> 444 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1)..(444) <220> <223> Nucleotide sequence of extracellular region of mouse FasL (Pro132-Leu279) <400> 147 ccc agt aca ccc tct gaa aaa aaa gag ccg agg agt gtg gcc cat tta 48 Pro Ser Thr Pro Ser Glu Lys Lys Glu Pro Arg Ser Val Ala His Leu 1 5 10 15 aca ggg aac ccc cac tca agg tcc atc cct ctg gaa tgg gaa gac aca 96 Thr Gly Asn Pro His Ser Arg Ser Ile Pro Leu Glu Trp Glu Asp Thr 20 25 30 tat gga acc gct ctg atc tct gga gtg aag tat aag aaa ggt ggc ctt 144 Tyr Gly Thr Ala Leu Ile Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 35 40 45 gtg atc aac gaa act ggg ttg tac ttc gtg tat tcc aaa gta tac ttc 192 Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe 50 55 60 cgg ggt cag tct tgc aac aac cag ccc cta aac cac aag gtc tat atg 240 Arg Gly Gln Ser Cys Asn Asn Gln Pro Leu Asn His Lys Val Tyr Met 65 70 75 80 agg aac tct aag tat cct gag gat ctg gtg cta atg gag gag aag agg 288 Arg Asn Ser Lys Tyr Pro Glu Asp Leu Val Leu Met Glu Glu Lys Arg 85 90 95 ttg aac tac tgc act act gga cag ata tgg gcc cac agc agc tac ctg 336 Leu Asn Tyr Cys Thr Thr Gly Gln Ile Trp Ala His Ser Ser Tyr Leu 100 105 110 ggg gca gta ttc aat ctt acc agt gct gac cat tta tat gtc aac ata 384 Gly Ala Val Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Ile 115 120 125 tct caa ctc tct ctg atc aat ttt gag gaa tct aag acc ttt ttc ggc 432 Ser Gln Leu Ser Leu Ile Asn Phe Glu Glu Ser Lys Thr Phe Phe Gly 130 135 140 tta tat aag ctc 444 Leu Tyr Lys Leu 145 <210> 148 <211> 148 <212> PRT <213> Mus musculus <220> <223> Amino acid sequence of extracellular region of mouse FasL (Pro132-Leu279) <400> 148 Pro Ser Thr Pro Ser Glu Lys Lys Glu Pro Arg Ser Val Ala His Leu 1 5 10 15 Thr Gly Asn Pro His Ser Arg Ser Ile Pro Leu Glu Trp Glu Asp Thr 20 25 30 Tyr Gly Thr Ala Leu Ile Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu 35 40 45 Val Ile Asn Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe 50 55 60 Arg Gly Gln Ser Cys Asn Asn Gln Pro Leu Asn His Lys Val Tyr Met 65 70 75 80 Arg Asn Ser Lys Tyr Pro Glu Asp Leu Val Leu Met Glu Glu Lys Arg 85 90 95 Leu Asn Tyr Cys Thr Thr Gly Gln Ile Trp Ala His Ser Ser Tyr Leu 100 105 110 Gly Ala Val Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Ile 115 120 125 Ser Gln Leu Ser Leu Ile Asn Phe Glu Glu Ser Lys Thr Phe Phe Gly 130 135 140 Leu Tyr Lys Leu 145 <210> 149 <211> 1212 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1212) <220> <223> Nucleotide sequence of Chimera A-Fc (g1S) <400> 149 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag ccc aaa tct 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc ctg 576 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc ctc 624 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg agc 672 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag 720 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc acg 768 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aat 816 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc ccc 864 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca cag 912 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag gtc 960 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc gtg 1008 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct 1056 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc acc 1104 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc gtg 1152 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc ctg 1200 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 tct ccg ggt aaa 1212 Ser Pro Gly Lys <210> 150 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (g1S) <400> 150 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 151 <211> 1293 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1293) <220> <223> Nucleotide sequence of OPG K194-Fc <400> 151 atg aac aac ttg ctg tgc tgc gcg ctc gtg ttt ctg gac atc tcc att 48 Met Asn Asn Leu Leu Cys Cys Ala Leu Val Phe Leu Asp Ile Ser Ile 1 5 10 15 aag tgg acc acc cag gaa acg ttt cct cca aag tac ctt cat tat gac 96 Lys Trp Thr Thr Gln Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp 20 25 30 gaa gaa acc tct cat cag ctg ttg tgt gac aaa tgt cct cct ggt acc 144 Glu Glu Thr Ser His Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr 35 40 45 tac cta aaa caa cac tgt aca gca aag tgg aag acc gtg tgc gcc cct 192 Tyr Leu Lys Gln His Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro 50 55 60 tgc cct gac cac tac tac aca gac agc tgg cac acc agt gac gag tgt 240 Cys Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys 65 70 75 80 cta tac tgc agc ccc gtg tgc aag gag ctg cag tac gtc aag cag gag 288 Leu Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu 85 90 95 tgc aat cgc acc cac aac cgc gtg tgc gaa tgc aag gaa ggg cgc tac 336 Cys Asn Arg Thr His Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr 100 105 110 ctt gag ata gag ttc tgc ttg aaa cat agg agc tgc cct cct gga ttt 384 Leu Glu Ile Glu Phe Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe 115 120 125 gga gtg gtg caa gct gga acc cca gag cga aat aca gtt tgc aaa aga 432 Gly Val Val Gln Ala Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg 130 135 140 tgt cca gat ggg ttc ttc tca aat gag acg tca tct aaa gca ccc tgt 480 Cys Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys 145 150 155 160 aga aaa cac aca aat tgc agt gtc ttt ggt ctc ctg cta act cag aaa 528 Arg Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys 165 170 175 gga aat gca aca cac gac aac ata tgt tcc gga aac agt gaa tca act 576 Gly Asn Ala Thr His Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr 180 185 190 caa aaa atc gag ggg cgc atg gat ccc aaa tct tcc gac aaa act cac 624 Gln Lys Ile Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His 195 200 205 aca tgc cca ccg tgc cca gca cct gaa ctc ctg ggg gga ccg tca gtc 672 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 210 215 220 ttc ctc ttc ccc cca aaa ccc aag gac acc ctc atg atc tcc cgg acc 720 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 225 230 235 240 cct gag gtc aca tgc gtg gtg gtg gac gtg agc cac gaa gac cct gag 768 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 245 250 255 gtc aag ttc aac tgg tac gtg gac ggc gtg gag gtg cat aat gcc aag 816 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 260 265 270 aca aag ccg cgg gag gag cag tac aac agc acg tac cgt gtg gtc agc 864 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 275 280 285 gtc ctc acc gtc ctg cac cag gac tgg ctg aat ggc aag gag tac aag 912 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 290 295 300 tgc aag gtc tcc aac aaa gcc ctc cca gcc ccc atc gag aaa acc atc 960 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 305 310 315 320 tcc aaa gcc aaa ggg cag ccc cga gaa cca cag gtg tac acc ctg ccc 1008 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 325 330 335 cca tcc cgg gat gag ctg acc aag aac cag gtc agc ctg acc tgc ctg 1056 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 340 345 350 gtc aaa ggc ttc tat ccc agc gac atc gcc gtg gag tgg gag agc aat 1104 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 355 360 365 ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg ctg gac tcc 1152 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 370 375 380 gac ggc tcc ttc ttc ctc tac agc aag ctc acc gtg gac aag agc agg 1200 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 385 390 395 400 tgg cag cag ggg aac gtc ttc tca tgc tcc gtg atg cat gag gct ctg 1248 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 405 410 415 cac aac cac tac acg cag aag agc ctc tcc ctg tct ccg ggt aaa 1293 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 420 425 430 <210> 152 <211> 431 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of OPG K194-Fc <400> 152 Met Asn Asn Leu Leu Cys Cys Ala Leu Val Phe Leu Asp Ile Ser Ile 1 5 10 15 Lys Trp Thr Thr Gln Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp 20 25 30 Glu Glu Thr Ser His Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr 35 40 45 Tyr Leu Lys Gln His Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro 50 55 60 Cys Pro Asp His Tyr Tyr Thr Asp Ser Trp His Thr Ser Asp Glu Cys 65 70 75 80 Leu Tyr Cys Ser Pro Val Cys Lys Glu Leu Gln Tyr Val Lys Gln Glu 85 90 95 Cys Asn Arg Thr His Asn Arg Val Cys Glu Cys Lys Glu Gly Arg Tyr 100 105 110 Leu Glu Ile Glu Phe Cys Leu Lys His Arg Ser Cys Pro Pro Gly Phe 115 120 125 Gly Val Val Gln Ala Gly Thr Pro Glu Arg Asn Thr Val Cys Lys Arg 130 135 140 Cys Pro Asp Gly Phe Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys 145 150 155 160 Arg Lys His Thr Asn Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys 165 170 175 Gly Asn Ala Thr His Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr 180 185 190 Gln Lys Ile Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His 195 200 205 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 210 215 220 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 225 230 235 240 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 245 250 255 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 260 265 270 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 275 280 285 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 290 295 300 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 305 310 315 320 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 325 330 335 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 340 345 350 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 355 360 365 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 370 375 380 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 385 390 395 400 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 405 410 415 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 420 425 430 <210> 153 <211> 330 <212> PRT <213> Homo sapiens <220> <221> PEPTIDE <222> (1)..(330) <220> <223> Amino acid sequence of heavy chain constant region of human IgG1 <400> 153 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 154 <211> 327 <212> PRT <213> Homo sapiens <220> <221> PEPTIDE <222> (1)..(327) <220> <223> Amino acid sequence of heavy chain constant region of human IgG4 <400> 154 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro 100 105 110 Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gly Gln Pro Arg 210 215 220 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly Lys 325 <210> 155 <211> 696 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(696) <220> <223> Nucleotide sequence of Fc (Eg1S) <400> 155 gag ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca 48 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc 96 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 aag gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg 144 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg 192 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag 240 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag 288 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc 336 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc 384 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc 432 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac 528 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac 576 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc 624 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 tca tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg cag aag 672 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 agc ctc tcc ctg tct ccg ggt aaa 696 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 156 <211> 232 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (Eg1S) <400> 156 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 157 <211> 696 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(696) <220> <223> Nucleotide sequence of Fc (Eg1S-YTE) <400> 157 gag ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca 48 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc 96 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 aag gac acc ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg 144 Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val 35 40 45 gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg 192 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag 240 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag 288 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc 336 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc 384 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc 432 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac 528 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac 576 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc 624 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 tca tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg cag aag 672 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 agc ctc tcc ctg tct ccg ggt aaa 696 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 158 <211> 232 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (Eg1S-YTE) <400> 158 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val 35 40 45 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 159 <211> 696 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(696) <220> <223> Nucleotide sequence of Fc (Eg1S-N434A) <400> 159 gag ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca 48 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc 96 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 aag gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg 144 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg 192 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag 240 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag 288 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc 336 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc 384 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc 432 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac 528 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac 576 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc 624 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 tca tgc tcc gtg atg cat gag gct ctg cac gcc cac tac acg cag aag 672 Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys 210 215 220 agc ctc tcc ctg tct ccg ggt aaa 696 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 160 <211> 232 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (Eg1S-N434A) <400> 160 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 161 <211> 693 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(693) <220> <223> Nucleotide sequence of Fc (g1S-LALAGA) <400> 161 ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct 48 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 gaa gcc gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag 96 Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg 144 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac 192 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac 240 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac 288 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc 336 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga 384 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag 432 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac 480 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag 528 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc 576 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca 624 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc 672 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 ctc tcc ctg tct ccg ggc aaa 693 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 162 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (g1S-LALAGA) <400> 162 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 163 <211> 696 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(696) <220> <223> Nucleotide sequence of Fc (Eg1S-LALAGA) <400> 163 gag ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca 48 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 cct gaa gcc gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc 96 Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 aag gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg 144 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg 192 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag 240 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag 288 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc 336 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc 384 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc 432 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac 528 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac 576 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc 624 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 tca tgc tcc gtg atg cat gag gct ctg cac aac cac tac acg cag aag 672 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 agc ctc tcc ctg tct ccg ggc aaa 696 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 164 <211> 232 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Fc (Eg1S-LALAGA) <400> 164 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 165 <211> 696 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(696) <220> <223> Nucleotide sequence of Fc (Eg1S-LALAGANA) <400> 165 gag ccc aaa tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca 48 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 cct gaa gcc gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc 96 Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 aag gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg 144 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg 192 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 gac ggc gtg gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag 240 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 tac aac agc acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag 288 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 gac tgg ctg aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc 336 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 ctc cca gcc ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc 384 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 cga gaa cca cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc 432 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 aag aac cag gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc 480 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 gac atc gcc gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac 528 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac 576 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 agc aag ctc acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc 624 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 tca tgc tcc gtg atg cat gag gct ctg cac gcc cac tac acg cag aag 672 Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys 210 215 220 agc ctc tcc ctg tct ccg ggt aaa 696 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 166 <211> 232 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (Eg1S-LALAGANA) <400> 166 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Lys Pro 20 25 30 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 167 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-Fc (Eg1S) <400> 167 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 168 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (Eg1S) <400> 168 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 169 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-Fc (Eg1S-YTE) <400> 169 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 170 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (Eg1S-YTE) <400> 170 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 171 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-Fc (Eg1S-N434A) <400> 171 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 172 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (Eg1S-N434A) <400> 172 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 173 <211> 1212 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1212) <220> <223> Nucleotide sequence of Chimera A-Fc (g1S-LALAGA) <400> 173 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag ccc aaa tct 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc gct 576 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc ctc 624 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg agc 672 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag 720 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc acg 768 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aat 816 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc ccc 864 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca cag 912 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag gtc 960 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc gtg 1008 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct 1056 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc acc 1104 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc gtg 1152 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc ctg 1200 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 tct ccg ggc aaa 1212 Ser Pro Gly Lys <210> 174 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (g1S-LALAGA) <400> 174 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 175 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-Fc (Eg1S-LALAGA) <400> 175 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 176 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (Eg1S-LALAGA) <400> 176 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 177 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-Fc (Eg1S-LALAGANA) <400> 177 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 178 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (Eg1S-LALAGANA) <400> 178 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 179 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57R <400> 179 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 180 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R <400> 180 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 181 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57V <400> 181 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 182 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V <400> 182 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 183 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D <400> 183 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 184 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D <400> 184 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 185 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E <400> 185 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 186 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E <400> 186 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 187 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D <400> 187 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 188 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D <400> 188 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 189 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57R-Fc (g4PEK) <400> 189 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 190 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (g4PEK) <400> 190 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 191 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57V-Fc (g4PEK) <400> 191 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 192 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (g4PEK) <400> 192 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 193 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D-Fc (g4PEK) <400> 193 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 194 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (g4PEK) <400> 194 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 195 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-Fc (g4PEK) <400> 195 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 196 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (g4PEK) <400> 196 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 197 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D-Fc (g4PEK) <400> 197 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 198 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (g4PEK) <400> 198 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 199 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K-Fc (Eg1S) <400> 199 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 200 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (Eg1S) <400> 200 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 201 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57L-Fc (Eg1S) <400> 201 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 202 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (Eg1S) <400> 202 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 203 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R-Fc (Eg1S) <400> 203 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 204 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (Eg1S) <400> 204 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 205 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57V-Fc (Eg1S) <400> 205 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 206 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (Eg1S) <400> 206 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 207 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D-Fc (Eg1S) <400> 207 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 208 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (Eg1S) <400> 208 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 209 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-Fc (Eg1S) <400> 209 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 210 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (Eg1S) <400> 210 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 211 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D-Fc (Eg1S) <400> 211 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 212 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (Eg1S) <400> 212 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 213 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K-Fc (Eg1S YTE) <400> 213 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 214 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (Eg1S YTE) <400> 214 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 215 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57L-Fc (Eg1S YTE) <400> 215 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 216 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (Eg1S YTE) <400> 216 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 217 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R-Fc (Eg1S YTE) <400> 217 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 218 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (Eg1S YTE) <400> 218 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 219 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57V-Fc (Eg1S YTE) <400> 219 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 220 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (Eg1S YTE) <400> 220 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 221 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D-Fc (Eg1S YTE) <400> 221 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 222 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (Eg1S YTE) <400> 222 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 223 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-Fc (Eg1S YTE) <400> 223 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 224 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (Eg1S YTE) <400> 224 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 225 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D-Fc (Eg1S YTE) <400> 225 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc tat atc act cgg gag cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 226 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (Eg1S YTE) <400> 226 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 227 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K-Fc (Eg1S N434A) <400> 227 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 228 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (Eg1S N434A) <400> 228 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 229 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57L-Fc (Eg1S N434A) <400> 229 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 230 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (Eg1S N434A) <400> 230 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 231 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R-Fc (Eg1S N434A) <400> 231 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 232 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (Eg1S N434A) <400> 232 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 233 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57V-Fc (Eg1S N434A) <400> 233 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 234 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (Eg1S N434A) <400> 234 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 235 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D-Fc (Eg1S N434A) <400> 235 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 236 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (Eg1S N434A) <400> 236 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 237 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-Fc (Eg1S N434A) <400> 237 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 238 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (Eg1S N434A) <400> 238 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 239 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D-Fc (Eg1S N434A) <400> 239 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa ctc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 240 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (Eg1S N434A) <400> 240 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 241 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K-Fc (Eg1S LALAGA) <400> 241 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 242 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (Eg1S LALAGA) <400> 242 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 243 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57L-Fc (Eg1S LALAGA) <400> 243 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 244 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (Eg1S LALAGA) <400> 244 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 245 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R-Fc (Eg1S LALAGA) <400> 245 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 246 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (Eg1S LALAGA) <400> 246 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 247 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57V-Fc (Eg1S LALAGA) <400> 247 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 248 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (Eg1S LALAGA) <400> 248 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 249 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D-Fc (Eg1S LALAGA) <400> 249 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 250 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (Eg1S LALAGA) <400> 250 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 251 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-Fc (Eg1S LALAGA) <400> 251 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 252 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (Eg1S LALAGA) <400> 252 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 253 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D-Fc (Eg1S LALAGA) <400> 253 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggc aaa 1215 Leu Ser Pro Gly Lys 405 <210> 254 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (Eg1S LALAGA) <400> 254 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 255 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K-Fc (Eg1S LALAGANA) <400> 255 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 256 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (Eg1S LALAGANA) <400> 256 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 257 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57L-Fc (Eg1S LALAGANA) <400> 257 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 258 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (Eg1S LALAGANA) <400> 258 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 259 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R-Fc (Eg1S LALAGANA) <400> 259 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 260 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (Eg1S LALAGANA) <400> 260 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 261 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57V-Fc (Eg1S LALAGANA) <400> 261 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 262 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (Eg1S LALAGANA) <400> 262 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 263 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58D-Fc (Eg1S LALAGANA) <400> 263 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 264 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (Eg1S LALAGANA) <400> 264 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 265 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-Fc (Eg1S LALAGANA) <400> 265 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 266 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (Eg1S LALAGANA) <400> 266 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 267 <211> 1215 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1215) <220> <223> Nucleotide sequence of Chimera A-E57R_R58D-Fc (Eg1S LALAGANA) <400> 267 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cgg gac tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag ccc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 tct tcc gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa gcc 576 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 gct ggg gcc ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag gac acc 624 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg gtg gac gtg 672 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 agc cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg 720 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 gag gtg cat aat gcc aag aca aag ccg cgg gag gag cag tac aac agc 768 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 acg tac cgt gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg 816 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 aat ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc ctc cca gcc 864 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 ccc atc gag aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gaa cca 912 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 cag gtg tac acc ctg ccc cca tcc cgg gat gag ctg acc aag aac cag 960 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 gtc agc ctg acc tgc ctg gtc aaa ggc ttc tat ccc agc gac atc gcc 1008 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 gtg gag tgg gag agc aat ggg cag ccg gag aac aac tac aag acc acg 1056 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc 1104 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 acc gtg gac aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc 1152 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 gtg atg cat gag gct ctg cac gcc cac tac acg cag aag agc ctc tcc 1200 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 ctg tct ccg ggt aaa 1215 Leu Ser Pro Gly Lys 405 <210> 268 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (Eg1S LALAGANA) <400> 268 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Pro Lys 165 170 175 Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala 180 185 190 Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys 405 <210> 269 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57A <400> 269 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gcc aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Ala Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 270 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57A <400> 270 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Ala Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 271 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57F <400> 271 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ttc aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Phe Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 272 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57F <400> 272 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Phe Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 273 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57H <400> 273 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cac aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu His Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 274 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57H <400> 274 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu His Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 275 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57I <400> 275 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg atc aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Ile Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 276 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57I <400> 276 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Ile Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 277 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57M <400> 277 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg atg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Met Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 278 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57M <400> 278 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Met Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 279 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57K_R58T <400> 279 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aca tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Thr Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 280 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58T <400> 280 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Thr Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 281 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57L_R58E <400> 281 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 282 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L_R58E <400> 282 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 283 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57V_R58T <400> 283 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aca tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Thr Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 284 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V_R58T <400> 284 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Thr Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 285 <211> 519 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(519) <220> <223> Nucleotide sequence of Chimera A-E57V_R58E <400> 285 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 286 <211> 173 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V_R58E <400> 286 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 287 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57A-Fc (g4PEK) <400> 287 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gcc aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Ala Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 288 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57A-Fc (g4PEK) <400> 288 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Ala Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 289 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57F-Fc (g4PEK) <400> 289 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ttc aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Phe Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 290 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57F-Fc (g4PEK) <400> 290 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Phe Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 291 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57H-Fc (g4PEK) <400> 291 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg cac aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu His Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 292 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57H-Fc (g4PEK) <400> 292 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu His Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 293 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57I-Fc (g4PEK) <400> 293 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg atc aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Ile Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 294 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57I-Fc (g4PEK) <400> 294 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Ile Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 295 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57M-Fc (g4PEK) <400> 295 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg atg aga tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Met Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 296 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57M-Fc (g4PEK) <400> 296 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Met Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 297 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57K_R58T-Fc (g4PEK) <400> 297 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg aag aca tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Thr Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 298 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58T-Fc (g4PEK) <400> 298 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Thr Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 299 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57L_R58E-Fc (g4PEK) <400> 299 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg ctg gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 300 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L_R58E-Fc (g4PEK) <400> 300 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 301 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57V_R58T-Fc (g4PEK) <400> 301 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg aca tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Thr Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 302 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V_R58T-Fc (g4PEK) <400> 302 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Thr Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 303 <211> 1206 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(1206) <220> <223> Nucleotide sequence of Chimera A-E57V_R58E-Fc (g4PEK) <400> 303 gag acc ttt ccc cct aag tac ctc cat tac gac gaa gaa aca agc cac 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 caa ctc ctc tgt gac aag tgc ccc cca ggg aca tat ttg aaa cag cac 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt acc gca aag tgg aaa aca gta tgc gcc cct tgc cca cca aga cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttt tgg aat tac ctg gtg gag tgc cgc tat tgt aac gtt 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggt gag cgt gag gaa gaa gct cga gca tgc cat gct act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgt gct tgc cgg tgt aga acc ggt ttt ttt gcc cat gca ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgc ctg gaa cat gca agc tgt ccc cct ggc gct ggc gtt atc gca ccc 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 gga act cct agc cag aat aca caa tgt cag cct tgc cct gac ggc ttt 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tct aat gag act tca agt aaa gct ccc tgc cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt agt gtg ttt ggg ttg ttg ctg acc cag aag ggg aac gca act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac att tgt agc ggg aat tca gaa agc acc caa aag gag tcc aaa 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 tat ggt ccc cca tgc cca cca tgc cca gca cct gag ttc gag ggg gga 576 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 cca tca gtc ttc ctg ttc ccc cca aaa ccc aag gac act ctc atg atc 624 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 tcc cgg acc cct gag gtc acg tgc gtg gtg gtg gac gtg agc cag gaa 672 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 gac ccc gag gtc cag ttc aac tgg tac gtg gat ggc gtg gag gtg cat 720 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 aat gcc aag aca aag ccg cgg gag gag cag ttc aac agc acg tac cgt 768 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 gtg gtc agc gtc ctc acc gtc ctg cac cag gac tgg ctg aac ggc aag 816 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 gag tac aag tgc aag gtc tcc aac aaa ggc ctc ccg tcc tcc atc gag 864 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 aaa acc atc tcc aaa gcc aaa ggg cag ccc cga gag cca cag gtg tac 912 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 acc ctg ccc cca tcc cag gag gag atg acc aag aac cag gtc agc ctg 960 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 acc tgc ctg gtc aaa ggc ttc tac ccc agc gac atc gcc gtg gag tgg 1008 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 gag agc aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg 1056 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 ctg gac tcc gac ggc tcc ttc ttc ctc tac agc aag cta acc gtg gac 1104 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 aag agc agg tgg cag gag ggg aat gtc ttc tca tgc tcc gtg atg cat 1152 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 gag gct ctg cac aac cac tac aca cag aag agc ctc tcc ctg tct ctg 1200 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 ggt aaa 1206 Gly Lys <210> 304 <211> 402 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V_R58E-Fc (g4PEK) <400> 304 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Glu Ser Lys 165 170 175 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly 180 185 190 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 195 200 205 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 210 215 220 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 225 230 235 240 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 245 250 255 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 260 265 270 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 275 280 285 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 290 295 300 Thr Leu Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 305 310 315 320 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 325 330 335 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 340 345 350 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 355 360 365 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 370 375 380 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 385 390 395 400 Gly Lys <210> 305 <211> 528 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(528) <220> <223> Nucleotide sequence of soluble recombinant of LIGHT with FLAG tag <400> 305 gac tac aaa gac gat gac gac aag ctt gac gga cct gca ggc tcc tgg 48 Asp Tyr Lys Asp Asp Asp Asp Lys Leu Asp Gly Pro Ala Gly Ser Trp 1 5 10 15 gag cag ctg ata caa gag cga agg tct cac gag gtc aac cca gca gcg 96 Glu Gln Leu Ile Gln Glu Arg Arg Ser His Glu Val Asn Pro Ala Ala 20 25 30 cat ctc aca ggg gcc aac tcc agc ttg acc ggc agc ggg ggg ccg ctg 144 His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu 35 40 45 tta tgg gag act cag ctg ggc ctg gcc ttc ctg agg ggc ctc agc tac 192 Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr 50 55 60 cac gat ggg gcc ctt gtg gtc acc aaa gct ggc tac tac tac atc tac 240 His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr 65 70 75 80 tcc aag gtg cag ctg ggc ggt gtg ggc tgc ccg ctg ggc ctg gcc agc 288 Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser 85 90 95 acc atc acc cac ggc ctc tac aag cgc aca ccc cgc tac ccc gag gag 336 Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu 100 105 110 ctg gag ctg ttg gtc agc cag cag tca ccc tgc gga cgg gcc acc agc 384 Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser 115 120 125 agc tcc cgg gtc tgg tgg gac agc agc ttc ctg ggt ggt gtg gta cac 432 Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His 130 135 140 ctg gag gct ggg gag aag gtg gtc gtc cgt gtg ctg gat gaa cgc ctg 480 Leu Glu Ala Gly Glu Lys Val Val Val Arg Val Leu Asp Glu Arg Leu 145 150 155 160 gtt cga ctg cgt gat ggt acc cgg tct tac ttc ggg gct ttc atg gtg 528 Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val 165 170 175 <210> 306 <211> 176 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of soluble recombinant of LIGHT with FLAG tag <400> 306 Asp Tyr Lys Asp Asp Asp Asp Lys Leu Asp Gly Pro Ala Gly Ser Trp 1 5 10 15 Glu Gln Leu Ile Gln Glu Arg Arg Ser His Glu Val Asn Pro Ala Ala 20 25 30 His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu 35 40 45 Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr 50 55 60 His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr 65 70 75 80 Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser 85 90 95 Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu 100 105 110 Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser 115 120 125 Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His 130 135 140 Leu Glu Ala Gly Glu Lys Val Val Val Arg Val Leu Asp Glu Arg Leu 145 150 155 160 Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val 165 170 175 <210> 307 <211> 777 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(777) <220> <223> Nucleotide sequence of TL1A, wherein the FLAG tag has been inserted into N-terminus thereof <400> 307 atg gac tac aag gac gac gat gac aag gcc gag gat ctg gga ctg agc 48 Met Asp Tyr Lys Asp Asp Asp Asp Lys Ala Glu Asp Leu Gly Leu Ser 1 5 10 15 ttt ggg gaa aca gcc agt gtg gaa atg ctg cca gag cac ggc agc tgc 96 Phe Gly Glu Thr Ala Ser Val Glu Met Leu Pro Glu His Gly Ser Cys 20 25 30 agg ccc aag gcc agg agc agc agc gca cgc tgg gct ctc acc tgc tgc 144 Arg Pro Lys Ala Arg Ser Ser Ser Ala Arg Trp Ala Leu Thr Cys Cys 35 40 45 ctg gtg ttg ctc ccc ttc ctt gca gga ctc acc aca tac ctg ctt gtc 192 Leu Val Leu Leu Pro Phe Leu Ala Gly Leu Thr Thr Tyr Leu Leu Val 50 55 60 agc cag ctc cgg gcc cag gga gag gcc tgt gtg cag ttc cag gct cta 240 Ser Gln Leu Arg Ala Gln Gly Glu Ala Cys Val Gln Phe Gln Ala Leu 65 70 75 80 aaa gga cag gag ttt gca cct tca cat cag caa gtt tat gca cct ctt 288 Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr Ala Pro Leu 85 90 95 aga gca gac gga gat aag cca agg gca cac ctg aca gtt gtg aga caa 336 Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val Val Arg Gln 100 105 110 act ccc aca cag cac ttt aaa aat cag ttc cca gct ctg cac tgg gaa 384 Thr Pro Thr Gln His Phe Lys Asn Gln Phe Pro Ala Leu His Trp Glu 115 120 125 cat gaa cta ggc ctg gcc ttc acc aag aac cga atg aac tat acc aac 432 His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn Tyr Thr Asn 130 135 140 aaa ttc ctg ctg atc cca gag tcg gga gac tac ttc att tac tcc cag 480 Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser Gln 145 150 155 160 gtc aca ttc cgt ggg atg acc tct gag tgc agt gaa atc aga caa gca 528 Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile Arg Gln Ala 165 170 175 ggc cga cca aac aag cca gac tcc atc act gtg gtc atc acc aag gta 576 Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile Thr Lys Val 180 185 190 aca gac agc tac cct gag cca acc cag ctc ctc atg ggg acc aag tct 624 Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly Thr Lys Ser 195 200 205 gta tgc gaa gta ggt agc aac tgg ttc cag ccc atc tac ctc gga gcc 672 Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr Leu Gly Ala 210 215 220 atg ttc tcc ttg caa gaa ggg gac aag cta atg gtg aac gtc agt gac 720 Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn Val Ser Asp 225 230 235 240 atc tct ttg gtg gat tac aca aaa gaa gat aaa acc ttc ttt gga gcc 768 Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly Ala 245 250 255 ttc tta cta 777 Phe Leu Leu <210> 308 <211> 259 <212> PRT <213> Homo sapiens <220> <223> The amino acid sequence of TL1A, wherein the FLAG tag has been inserted into N-terminus thereof <400> 308 Met Asp Tyr Lys Asp Asp Asp Asp Lys Ala Glu Asp Leu Gly Leu Ser 1 5 10 15 Phe Gly Glu Thr Ala Ser Val Glu Met Leu Pro Glu His Gly Ser Cys 20 25 30 Arg Pro Lys Ala Arg Ser Ser Ser Ala Arg Trp Ala Leu Thr Cys Cys 35 40 45 Leu Val Leu Leu Pro Phe Leu Ala Gly Leu Thr Thr Tyr Leu Leu Val 50 55 60 Ser Gln Leu Arg Ala Gln Gly Glu Ala Cys Val Gln Phe Gln Ala Leu 65 70 75 80 Lys Gly Gln Glu Phe Ala Pro Ser His Gln Gln Val Tyr Ala Pro Leu 85 90 95 Arg Ala Asp Gly Asp Lys Pro Arg Ala His Leu Thr Val Val Arg Gln 100 105 110 Thr Pro Thr Gln His Phe Lys Asn Gln Phe Pro Ala Leu His Trp Glu 115 120 125 His Glu Leu Gly Leu Ala Phe Thr Lys Asn Arg Met Asn Tyr Thr Asn 130 135 140 Lys Phe Leu Leu Ile Pro Glu Ser Gly Asp Tyr Phe Ile Tyr Ser Gln 145 150 155 160 Val Thr Phe Arg Gly Met Thr Ser Glu Cys Ser Glu Ile Arg Gln Ala 165 170 175 Gly Arg Pro Asn Lys Pro Asp Ser Ile Thr Val Val Ile Thr Lys Val 180 185 190 Thr Asp Ser Tyr Pro Glu Pro Thr Gln Leu Leu Met Gly Thr Lys Ser 195 200 205 Val Cys Glu Val Gly Ser Asn Trp Phe Gln Pro Ile Tyr Leu Gly Ala 210 215 220 Met Phe Ser Leu Gln Glu Gly Asp Lys Leu Met Val Asn Val Ser Asp 225 230 235 240 Ile Ser Leu Val Asp Tyr Thr Lys Glu Asp Lys Thr Phe Phe Gly Ala 245 250 255 Phe Leu Leu <210> 309 <211> 867 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1)..(867) <220> <223> Nucleotide sequence of FasL wherein FLAG tag has been inserted into N-terminus thereof <400> 309 atg gac tac aag gac gac gat gac aag cag cag ccc ttc aat tac cca 48 Met Asp Tyr Lys Asp Asp Asp Asp Lys Gln Gln Pro Phe Asn Tyr Pro 1 5 10 15 tat ccc cag atc tac tgg gtg gac agc agt gcc agc tct ccc tgg gcc 96 Tyr Pro Gln Ile Tyr Trp Val Asp Ser Ser Ala Ser Ser Pro Trp Ala 20 25 30 cct cca ggc aca gtt ctt ccc tgt cca acc tct gtg ccc aga agg cct 144 Pro Pro Gly Thr Val Leu Pro Cys Pro Thr Ser Val Pro Arg Arg Pro 35 40 45 ggt caa agg agg cca cca cca cca ccg cca ccg cca cca cta cca cct 192 Gly Gln Arg Arg Pro Pro Pro Pro Pro Pro Pro Pro Leu Pro Pro 50 55 60 ccg ccg ccg ccg cca cca ctg cct cca cta ccg ctg cca ccc ctg aag 240 Pro Pro Pro Pro Pro Pro Leu Pro Pro Leu Pro Leu Pro Pro Leu Lys 65 70 75 80 aag aga ggg aac cac agc aca ggc ctg tgt ctc ctt gtg atg ttt ttc 288 Lys Arg Gly Asn His Ser Thr Gly Leu Cys Leu Leu Val Met Phe Phe 85 90 95 atg gtt ctg gtt gcc ttg gta gga ttg ggc ctg ggg atg ttt cag ctc 336 Met Val Leu Val Ala Leu Val Gly Leu Gly Leu Gly Met Phe Gln Leu 100 105 110 ttc cac cta cag aag gag ctg gca gaa ctc cga gag tct acc agc cag 384 Phe His Leu Gln Lys Glu Leu Ala Glu Leu Arg Glu Ser Thr Ser Gln 115 120 125 atg cac aca gca tca tct ttg gag aag caa ata ggc cac ccc agt cca 432 Met His Thr Ala Ser Ser Leu Glu Lys Gln Ile Gly His Pro Ser Pro 130 135 140 ccc cct gaa aaa aag gag ctg agg aaa gtg gcc cat tta aca ggc aag 480 Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His Leu Thr Gly Lys 145 150 155 160 tcc aac tca agg tcc atg cct ctg gaa tgg gaa gac acc tat gga att 528 Ser Asn Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile 165 170 175 gtc ctg ctt tct gga gtg aag tat aag aag ggt ggc ctt gtg atc aat 576 Val Leu Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu Val Ile Asn 180 185 190 gaa act ggg ctg tac ttt gta tat tcc aaa gta tac ttc cgg ggt caa 624 Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe Arg Gly Gln 195 200 205 tct tgc aac aac ctg ccc ctg agc cac aag gtc tac atg agg aac tct 672 Ser Cys Asn Asn Leu Pro Leu Ser His Lys Val Tyr Met Arg Asn Ser 210 215 220 aag tat ccc cag gat ctg gtg atg atg gag ggg aag atg atg agc tac 720 Lys Tyr Pro Gln Asp Leu Val Met Met Glu Gly Lys Met Met Ser Tyr 225 230 235 240 tgc act act ggg cag atg tgg gcc cgc agc agc tac ctg ggg gca gtg 768 Cys Thr Thr Gly Gln Met Trp Ala Arg Ser Ser Tyr Leu Gly Ala Val 245 250 255 ttc aat ctt acc agt gct gat cat tta tat gtc aac gta tct gag ctc 816 Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Val Ser Glu Leu 260 265 270 tct ctg gtc aat ttt gag gaa tct cag acg ttt ttc ggc tta tat aag 864 Ser Leu Val Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly Leu Tyr Lys 275 280 285 ctc 867 Leu <210> 310 <211> 289 <212> PRT <213> Homo sapiens <220> <223> Amino acid sequence of FasL wherein FLAG tag has been inserted into N-terminus thereof <400> 310 Met Asp Tyr Lys Asp Asp Asp Asp Lys Gln Gln Pro Phe Asn Tyr Pro 1 5 10 15 Tyr Pro Gln Ile Tyr Trp Val Asp Ser Ser Ala Ser Ser Pro Trp Ala 20 25 30 Pro Pro Gly Thr Val Leu Pro Cys Pro Thr Ser Val Pro Arg Arg Pro 35 40 45 Gly Gln Arg Arg Pro Pro Pro Pro Pro Pro Pro Pro Leu Pro Pro 50 55 60 Pro Pro Pro Pro Pro Pro Leu Pro Pro Leu Pro Leu Pro Pro Leu Lys 65 70 75 80 Lys Arg Gly Asn His Ser Thr Gly Leu Cys Leu Leu Val Met Phe Phe 85 90 95 Met Val Leu Val Ala Leu Val Gly Leu Gly Leu Gly Met Phe Gln Leu 100 105 110 Phe His Leu Gln Lys Glu Leu Ala Glu Leu Arg Glu Ser Thr Ser Gln 115 120 125 Met His Thr Ala Ser Ser Leu Glu Lys Gln Ile Gly His Pro Ser Pro 130 135 140 Pro Pro Glu Lys Lys Glu Leu Arg Lys Val Ala His Leu Thr Gly Lys 145 150 155 160 Ser Asn Ser Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile 165 170 175 Val Leu Leu Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu Val Ile Asn 180 185 190 Glu Thr Gly Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe Arg Gly Gln 195 200 205 Ser Cys Asn Asn Leu Pro Leu Ser His Lys Val Tyr Met Arg Asn Ser 210 215 220 Lys Tyr Pro Gln Asp Leu Val Met Met Glu Gly Lys Met Met Ser Tyr 225 230 235 240 Cys Thr Thr Gly Gln Met Trp Ala Arg Ser Ser Tyr Leu Gly Ala Val 245 250 255 Phe Asn Leu Thr Ser Ala Asp His Leu Tyr Val Asn Val Ser Glu Leu 260 265 270 Ser Leu Val Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly Leu Tyr Lys 275 280 285 Leu <210> 311 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (g1S YTE) <400> 311 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro Lys 20 25 30 Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 312 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (g1S N434A) <400> 312 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 313 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (g1S LALAGANA) <400> 313 Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 1 5 10 15 Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 314 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (g1S YTE) <400> 314 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 315 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (g1S N434A) <400> 315 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 316 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-Fc (g1S LALAGANA) <400> 316 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 317 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (g1S YTE) <400> 317 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 318 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (g1S YTE) <400> 318 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 319 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (g1S YTE) <400> 319 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 320 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (g1S YTE) <400> 320 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 321 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (g1S YTE) <400> 321 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 322 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (g1S YTE) <400> 322 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 323 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (g1S YTE) <400> 323 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 324 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (g1S N434A) <400> 324 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 325 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (g1S N434A) <400> 325 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 326 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (g1S N434A) <400> 326 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 327 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (g1S N434A) <400> 327 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 328 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (g1S N434A) <400> 328 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 329 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (g1S N434A) <400> 329 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 330 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (g1S N434A) <400> 330 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 331 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K-Fc (g1S LALAGANA) <400> 331 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 332 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57L-Fc (g1S LALAGANA) <400> 332 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Leu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 333 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R-Fc (g1S LALAGANA) <400> 333 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 334 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57V-Fc (g1S LALAGANA) <400> 334 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Val Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 335 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58D-Fc (g1S LALAGANA) <400> 335 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 336 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57K_R58E-Fc (g1S LALAGANA) <400> 336 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 337 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Amino acid sequence of Chimera A-E57R_R58D-Fc (g1S LALAGANA) <400> 337 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Arg Asp Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys Pro Lys Ser 165 170 175 Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 180 185 190 Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys <210> 338 <211> 711 <212> DNA <213> Artificial Sequence <220> <221> CDS <222> (1)..(711) <220> <223> Nucleotide sequence of Fc (IEGRMD g1S) <400> 338 atc gag ggg cgc atg gat ccc aaa tct tcc gac aaa act cac aca tgc 48 Ile Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys 1 5 10 15 cca ccg tgc cca gca cct gaa ctc ctg ggg gga ccg tca gtc ttc ctc 96 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 20 25 30 ttc ccc cca aaa ccc aag gac acc ctc atg atc tcc cgg acc cct gag 144 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 35 40 45 gtc aca tgc gtg gtg gtg gac gtg agc cac gaa gac cct gag gtc aag 192 Val Thr Cys Val Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 50 55 60 ttc aac tgg tac gtg gac ggc gtg gag gtg cat aat gcc aag aca aag 240 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 65 70 75 80 ccg cgg gag gag cag tac aac agc acg tac cgt gtg gtc agc gtc ctc 288 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 85 90 95 acc gtc ctg cac cag gac tgg ctg aat ggc aag gag tac aag tgc aag 336 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 100 105 110 gtc tcc aac aaa gcc ctc cca gcc ccc atc gag aaa acc atc tcc aaa 384 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 115 120 125 gcc aaa ggg cag ccc cga gaa cca cag gtg tac acc ctg ccc cca tcc 432 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 130 135 140 cgg gat gag ctg acc aag aac cag gtc agc ctg acc tgc ctg gtc aaa 480 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 145 150 155 160 ggc ttc tat ccc agc gac atc gcc gtg gag tgg gag agc aat ggg cag 528 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 165 170 175 ccg gag aac aac tac aag acc acg cct ccc gtg ctg gac tcc gac ggc 576 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 180 185 190 tcc ttc ttc ctc tac agc aag ctc acc gtg gac aag agc agg tgg cag 624 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 195 200 205 cag ggg aac gtc ttc tca tgc tcc gtg atg cat gag gct ctg cac aac 672 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 210 215 220 cac tac acg cag aag agc ctc tcc ctg tct ccg ggt aaa 711 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 225 230 235 <210> 339 <211> 237 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Fc (IEGRMD g1S) <400> 339 Ile Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys 1 5 10 15 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 20 25 30 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 35 40 45 Val Thr Cys Val Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 50 55 60 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 65 70 75 80 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 85 90 95 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 100 105 110 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 115 120 125 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 130 135 140 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 145 150 155 160 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 165 170 175 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 180 185 190 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 195 200 205 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 210 215 220 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 225 230 235 <210> 340 <211> 508 <212> PRT <213> Artifical Sequence <220> <223> amino acid sequence of R218Q-Fc <400> 340 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser Cys Thr Gly Phe Pro Leu Ser Thr Arg Val 165 170 175 Pro Gly Ala Glu Glu Cys Glu Arg Ala Val Ile Asp Phe Val Ala Phe 180 185 190 Gln Asp Ile Ser Ile Lys Arg Leu Gln Arg Leu Leu Gln Ala Leu Glu 195 200 205 Ala Pro Glu Gly Trp Gly Pro Thr Pro Gln Ala Gly Arg Ala Ala Leu 210 215 220 Gln Leu Lys Leu Arg Arg Arg Leu Thr Glu Leu Leu Gly Ala Gln Asp 225 230 235 240 Gly Ala Leu Leu Val Arg Leu Leu Gln Ala Leu Arg Val Ala Arg Met 245 250 255 Pro Gly Leu Glu Arg Ser Val Arg Glu Arg Phe Leu Pro Val His Ile 260 265 270 Glu Gly Arg Met Asp Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro 275 280 285 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 290 295 300 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 305 310 315 320 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 325 330 335 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 340 345 350 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 355 360 365 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 370 375 380 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 385 390 395 400 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 405 410 415 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 420 425 430 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 435 440 445 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 450 455 460 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 465 470 475 480 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 485 490 495 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 500 505 <210> 341 <211> 528 <212> DNA <213> <220> <221> CDS <222> (1)..(528) <220> <223> Nucleotide sequence of FLAG-cynoLIGHT <400> 341 gac tac aaa gac gat gac gac aag gac gga cac gga ggc tcc tgg gag 48 Asp Tyr Lys Asp Asp Asp Asp Lys Asp Gly His Gly Gly Ser Trp Glu 1 5 10 15 cag ctg ata caa gag cga agg tct cac cag aat aac cca gca gca cat 96 Gln Leu Ile Gln Glu Arg Arg Ser His Gln Asn Asn Pro Ala Ala His 20 25 30 ctc aca ggg gcc aac tcc agc ttg acg ggc agc ggg gga ccg ctg cta 144 Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu 35 40 45 tgg gag acc cag ctg ggc ctg gcc ttc ctg agg ggc ctc gac tac cac 192 Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Asp Tyr His 50 55 60 gat ggg gcc ctt gtg gcc aac aag gct ggc tac tac tac atc tac tcc 240 Asp Gly Ala Leu Val Ala Asn Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser 65 70 75 80 aag gtg cag ctg ggc ggt gtg ggc tgc ccg ctg ggc ctg gcc agc acc 288 Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr 85 90 95 atc acc cac ggc ctc tac aag cgc acg ccc cgc tac ccc gag gag ctg 336 Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu 100 105 110 gag ctg ttg gtc agc cag cag tca ccc tgc gga cgg gcc acc agc aac 384 Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Asn 115 120 125 tcc cgc gtc tgg tgg gac agc agc ttc ctg ggt ggt gtg gtg cac ctg 432 Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu 130 135 140 gag gtt ggg gag gag gtg gtc gtc cgt gtg ctg gat gag cgc ctg gtc 480 Glu Val Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val 145 150 155 160 cga ctg cgt gat ggc acc cgg tct tac ttc ggg gct ttc atg gtg tga 528 Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val 165 170 175 <210> 342 <211> 175 <212> PRT <213> <220> <223> amino acid sequence of FLAG-cynoLIGHT <400> 342 Asp Tyr Lys Asp Asp Asp Asp Lys Asp Gly His Gly Gly Ser Trp Glu 1 5 10 15 Gln Leu Ile Gln Glu Arg Arg Ser His Gln Asn Asn Pro Ala Ala His 20 25 30 Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu 35 40 45 Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Asp Tyr His 50 55 60 Asp Gly Ala Leu Val Ala Asn Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser 65 70 75 80 Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr 85 90 95 Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu 100 105 110 Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Asn 115 120 125 Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu 130 135 140 Glu Val Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val 145 150 155 160 Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val 165 170 175 <210> 343 <211> 666 <212> DNA <213> <220> <221> CDS <222> (1)..(666) <220> <223> Nucleotide sequence of a synthetic construct obtained by deleting a cleavage site and an intracellular proline-rich domain from FasL and insert a FLAG tag into N-terminus of FasL <400> 343 gac tac aag gac gac gat gac aag cag cag ccc ttc aat tac ccc ctg 48 Asp Tyr Lys Asp Asp Asp Asp Lys Gln Gln Pro Phe Asn Tyr Pro Leu 1 5 10 15 aag aag aga ggg aac cac agc aca ggc ctg tgt ctc ctt gtg atg ttt 96 Lys Lys Arg Gly Asn His Ser Thr Gly Leu Cys Leu Leu Val Met Phe 20 25 30 ttc atg gtt ctg gtt gcc ttg gta gga ttg ggc ctg ggg atg ttt cag 144 Phe Met Val Leu Val Ala Leu Val Gly Leu Gly Leu Gly Met Phe Gln 35 40 45 ctc ttc cac cta cag aag gag ctg gca gaa ctc cga gag tct acc agc 192 Leu Phe His Leu Gln Lys Glu Leu Ala Glu Leu Arg Glu Ser Thr Ser 50 55 60 cag atg cac aca gca tca tct ttg ggc cac ccc agt cca ccc cct gaa 240 Gln Met His Thr Ala Ser Ser Leu Gly His Pro Ser Pro Pro Pro Glu 65 70 75 80 aaa aag gag ctg agg aaa gtg gcc cat tta aca ggc aag tcc aac tca 288 Lys Lys Glu Leu Arg Lys Val Ala His Leu Thr Gly Lys Ser Asn Ser 85 90 95 agg tcc atg cct ctg gaa tgg gaa gac acc tat gga att gtc ctg ctt 336 Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu Leu 100 105 110 tct gga gtg aag tat aag aag ggt ggc ctt gtg atc aat gaa act ggg 384 Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly 115 120 125 ctg tac ttt gta tat tcc aaa gta tac ttc cgg ggt caa tct tgc aac 432 Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Asn 130 135 140 aac ctg ccc ctg agc cac aag gtc tac atg agg aac tct aag tat ccc 480 Asn Leu Pro Leu Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro 145 150 155 160 cag gat ctg gtg atg atg gag ggg aag atg atg agc tac tgc act act 528 Gln Asp Leu Val Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr 165 170 175 ggg cag atg tgg gcc cgc agc agc tac ctg ggg gca gtg ttc aat ctt 576 Gly Gln Met Trp Ala Arg Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu 180 185 190 acc agt gct gat cat tta tat gtc aac gta tct gag ctc tct ctg gtc 624 Thr Ser Ala Asp His Leu Tyr Val Asn Val Ser Glu Leu Ser Leu Val 195 200 205 aat ttt gag gaa tct cag acg ttt ttc ggc tta tat aag ctc 666 Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly Leu Tyr Lys Leu 210 215 220 <210> 344 <211> 222 <212> PRT <213> <220> <223> Amino acid sequence of a synthetic construct obtained by deleting a cleavage site and an intracellular proline-rich domain from FasL and insert a FLAG tag into N-terminus of FasL <400> 344 Asp Tyr Lys Asp Asp Asp Asp Lys Gln Gln Pro Phe Asn Tyr Pro Leu 1 5 10 15 Lys Lys Arg Gly Asn His Ser Thr Gly Leu Cys Leu Leu Val Met Phe 20 25 30 Phe Met Val Leu Val Ala Leu Val Gly Leu Gly Leu Gly Met Phe Gln 35 40 45 Leu Phe His Leu Gln Lys Glu Leu Ala Glu Leu Arg Glu Ser Thr Ser 50 55 60 Gln Met His Thr Ala Ser Ser Leu Gly His Pro Ser Pro Pro Pro Glu 65 70 75 80 Lys Lys Glu Leu Arg Lys Val Ala His Leu Thr Gly Lys Ser Asn Ser 85 90 95 Arg Ser Met Pro Leu Glu Trp Glu Asp Thr Tyr Gly Ile Val Leu Leu 100 105 110 Ser Gly Val Lys Tyr Lys Lys Gly Gly Leu Val Ile Asn Glu Thr Gly 115 120 125 Leu Tyr Phe Val Tyr Ser Lys Val Tyr Phe Arg Gly Gln Ser Cys Asn 130 135 140 Asn Leu Pro Leu Ser His Lys Val Tyr Met Arg Asn Ser Lys Tyr Pro 145 150 155 160 Gln Asp Leu Val Met Met Glu Gly Lys Met Met Ser Tyr Cys Thr Thr 165 170 175 Gly Gln Met Trp Ala Arg Ser Ser Tyr Leu Gly Ala Val Phe Asn Leu 180 185 190 Thr Ser Ala Asp His Leu Tyr Val Asn Val Ser Glu Leu Ser Leu Val 195 200 205 Asn Phe Glu Glu Ser Gln Thr Phe Phe Gly Leu Tyr Lys Leu 210 215 220 <210> 345 <211> 516 <212> DNA <213> <220> <221> CDS <222> (1)..(516) <220> <223> Nucleotide sequence of S195-His6 <400> 345 gta gca gag aca ccc acc tat cca tgg agg gac gct gag acc ggt gag 48 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 cgg ctc gtt tgc gcc caa tgt cca cca ggg acc ttc gtt cag cgg ccc 96 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 tgc cga cgg gac agt cca aca aca tgt ggc cct tgt cct ccc cgc cac 144 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg gag cgc tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgc cct ccc ggc act 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 ttt tct gcc tct tcc agt tca tca gag caa tgc cag cct cac aga aac 432 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 tgt acc gct ctg ggg ctc gcc ctg aat gtc ccc ggc tct tca agt cac 480 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 gat acc ctg tgc acc agc cat cat cat cat cat cat 516 Asp Thr Leu Cys Thr Ser His His His His His His 165 170 <210> 346 <211> 172 <212> PRT <213> <220> <223> amino acid sequence of S195-His6 <400> 346 Val Ala Glu Thr Pro Thr Tyr Pro Trp Arg Asp Ala Glu Thr Gly Glu 1 5 10 15 Arg Leu Val Cys Ala Gln Cys Pro Pro Gly Thr Phe Val Gln Arg Pro 20 25 30 Cys Arg Arg Asp Ser Pro Thr Thr Cys Gly Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Pro Gly Thr 115 120 125 Phe Ser Ala Ser Ser Ser Ser Ser Ser Glu Gln Cys Gln Pro His Arg Asn 130 135 140 Cys Thr Ala Leu Gly Leu Ala Leu Asn Val Pro Gly Ser Ser Ser His 145 150 155 160 Asp Thr Leu Cys Thr Ser His His His His His His 165 170 <210> 347 <211> 537 <212> DNA <213> <220> <221> CDS <222> (1)..(537) <220> <223> Nucleotide sequence of Chimera A-His6 <400> 347 gag act ttc ccc ccc aag tat ctg cac tat gat gag gag act tcc cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ctg tgt gac aag tgt ccc ccc ggg acc tat ctg aag cag cat 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt act gcc aag tgg aag act gtg tgt gct cct tgt cct ccc cgc cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg gag cgc tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgt ccc gat ggc ttc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tcc aat gag act tcc tcc aag gcc ccc tgt cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt tcc gtg ttc ggg ctg ctg ctg act cag aag ggg aat gcc act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac atc tgt tcc ggg aac tcc gag tcc act cag aag cat cat cat 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys His His His 165 170 175 cat cat cat 537 His His His <210> 348 <211> 179 <212> PRT <213> <220> <223> Amino acid sequence of Chimera A-His6 <400> 348 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys His His His 165 170 175 His His His <210> 349 <211> 537 <212> DNA <213> <220> <221> CDS <222> (1)..(537) <220> <223> Nucleotide sequence of Chimera A-E57K-His6 <400> 349 gag act ttc ccc ccc aag tat ctg cac tat gat gag gag act tcc cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ctg tgt gac aag tgt ccc ccc ggg acc tat ctg aag cag cat 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt act gcc aag tgg aag act gtg tgt gct cct tgt cct ccc cgc cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg aag cgc tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgt ccc gat ggc ttc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tcc aat gag act tcc tcc aag gcc ccc tgt cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt tcc gtg ttc ggg ctg ctg ctg act cag aag ggg aat gcc act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac atc tgt tcc ggg aac tcc gag tcc act cag aag cat cat cat 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys His His His 165 170 175 cat cat cat 537 His His His <210> 350 <211> 179 <212> PRT <213> <220> <223> Amino acid sequence of Chimera A-E57K-His6 <400> 350 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys His His His 165 170 175 His His His <210> 351 <211> 537 <212> DNA <213> <220> <221> CDS <222> (1)..(537) <220> <223> Nucleotide sequence of Chimera A-E57K_R58E-His6 <400> 351 gag act ttc ccc ccc aag tat ctg cac tat gat gag gag act tcc cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ctg tgt gac aag tgt ccc ccc ggg acc tat ctg aag cag cat 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt act gcc aag tgg aag act gtg tgt gct cct tgt cct ccc cgc cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg aag gag tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgt ccc gat ggc ttc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tcc aat gag act tcc tcc aag gcc ccc tgt cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt tcc gtg ttc ggg ctg ctg ctg act cag aag ggg aat gcc act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac atc tgt tcc ggg aac tcc gag tcc act cag aag cat cat cat 528 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys His His His 165 170 175 cat cat cat 537 His His His <210> 352 <211> 179 <212> PRT <213> <220> <223> Amino acid sequence of Chimera A-E57K_R58E-His6 <400> 352 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys His His His 165 170 175 His His His <210> 353 <211> 519 <212> DNA <213> <220> <221> CDS <222> (1)..(519) <220> <223> Optimized nucleotide sequence of Chimera A <400> 353 gag act ttc ccc ccc aag tat ctg cac tat gat gag gag act tcc cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ctg tgt gac aag tgt ccc ccc ggg acc tat ctg aag cag cat 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt act gcc aag tgg aag act gtg tgt gct cct tgt cct ccc cgc cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg gag cgc tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Glu Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgt ccc gat ggc ttc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tcc aat gag act tcc tcc aag gcc ccc tgt cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt tcc gtg ttc ggg ctg ctg ctg act cag aag ggg aat gcc act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac atc tgt tcc ggg aac tcc gag tcc act cag aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 354 <211> 519 <212> DNA <213> <220> <221> CDS <222> (1)..(519) <220> <223> Optimized nucleotide sequence of Chimera A-E57K <400> 354 gag act ttc ccc ccc aag tat ctg cac tat gat gag gag act tcc cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ctg tgt gac aag tgt ccc ccc ggg acc tat ctg aag cag cat 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt act gcc aag tgg aag act gtg tgt gct cct tgt cct ccc cgc cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg aag cgc tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Arg Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgt ccc gat ggc ttc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tcc aat gag act tcc tcc aag gcc ccc tgt cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt tcc gtg ttc ggg ctg ctg ctg act cag aag ggg aat gcc act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac atc tgt tcc ggg aac tcc gag tcc act cag aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170 <210> 355 <211> 519 <212> DNA <213> <220> <221> CDS <222> (1)..(519) <220> <223> Optimized nucleotide sequence of Chimera A-E57K_R58E <400> 355 gag act ttc ccc ccc aag tat ctg cac tat gat gag gag act tcc cat 48 Glu Thr Phe Pro Lys Tyr Leu His Tyr Asp Glu Glu Thr Ser His 1 5 10 15 cag ctg ctg tgt gac aag tgt ccc ccc ggg acc tat ctg aag cag cat 96 Gln Leu Leu Cys Asp Lys Cys Pro Pro Gly Thr Tyr Leu Lys Gln His 20 25 30 tgt act gcc aag tgg aag act gtg tgt gct cct tgt cct ccc cgc cac 144 Cys Thr Ala Lys Trp Lys Thr Val Cys Ala Pro Cys Pro Pro Arg His 35 40 45 tac act cag ttc tgg aac tac ctg aag gag tgt cgc tac tgc aat gtg 192 Tyr Thr Gln Phe Trp Asn Tyr Leu Lys Glu Cys Arg Tyr Cys Asn Val 50 55 60 ctg tgt ggg gaa cgc gag gag gag gct cgc gct tgt cat gcc act cac 240 Leu Cys Gly Glu Arg Glu Glu Glu Ala Arg Ala Cys His Ala Thr His 65 70 75 80 aat cgc gcc tgt cgc tgt cgc act ggc ttc ttc gcc cat gcc ggc ttc 288 Asn Arg Ala Cys Arg Cys Arg Thr Gly Phe Phe Ala His Ala Gly Phe 85 90 95 tgt ctg gag cat gcc tcc tgt cct cct ggg gct ggc gtg atc gct cct 336 Cys Leu Glu His Ala Ser Cys Pro Pro Gly Ala Gly Val Ile Ala Pro 100 105 110 ggg act ccc tcc cag aac act cag tgt cag ccc tgt ccc gat ggc ttc 384 Gly Thr Pro Ser Gln Asn Thr Gln Cys Gln Pro Cys Pro Asp Gly Phe 115 120 125 ttc tcc aat gag act tcc tcc aag gcc ccc tgt cgc aag cac acc aac 432 Phe Ser Asn Glu Thr Ser Ser Lys Ala Pro Cys Arg Lys His Thr Asn 130 135 140 tgt tcc gtg ttc ggg ctg ctg ctg act cag aag ggg aat gcc act cac 480 Cys Ser Val Phe Gly Leu Leu Leu Thr Gln Lys Gly Asn Ala Thr His 145 150 155 160 gac aac atc tgt tcc ggg aac tcc gag tcc act cag aag 519 Asp Asn Ile Cys Ser Gly Asn Ser Glu Ser Thr Gln Lys 165 170

Claims (49)

Wild-type decoy receptor (Decoy Receptor) 3 (hereinafter, abbreviated as DcR3) as a variant DcR3 variant, showing improved in vivo dynamics than the wild-type DcR3, the DcR3 variant. According to claim 1, DcR3 variant having one or more N- glycosidic linkage complex sugar chains. According to claim 1, LIGHT, TL1A, and DcR3 variant having neutralizing activity with respect to at least one of FasL. According to claim 1, LIGHT, TL1A and DcR3 variant having neutralizing activity against all of FasL. The DcR3 variant according to claim 1, which has no neutralizing activity with respect to FasL, and also has neutralizing activity against any one or more of LIGHT and TL1A. The DcR3 variant according to claim 1, which has no neutralizing activity with respect to FasL, and also has neutralizing activity against LIGHT and TL1A. In the cysteine-rich region of wild-type DcR3, at least a portion of the cysteine-rich domain (hereinafter abbreviated as CRD) of wild-type DcR3 is substituted with at least a portion of the cysteine-rich domain of a TNF receptor superfamily molecule other than DcR3. It contains an amino acid sequence a first chimeric cysteine-rich region or a second chimeric cysteine-rich region comprising an amino acid sequence in which 1 to 30 amino acids are deleted, substituted, inserted or added in the amino acid sequence of the first chimeric cysteine-rich region; DcR3 variant comprising. According to claim 7, DcR3 variant having one or more N- glycosidic linkage complex sugar chains. According to claim 7, LIGHT, TL1A, and DcR3 variant having neutralizing activity with respect to at least one of FasL. According to claim 7, LIGHT, TL1A and DcR3 variant having neutralizing activity against all of FasL. The DcR3 variant according to claim 7, which has no neutralizing activity with respect to FasL, and also has neutralizing activity against any one or more of LIGHT and TL1A. The DcR3 variant according to claim 7, which has no neutralizing activity with respect to FasL, and also has neutralizing activity against LIGHT and TL1A. The DcR3 variant according to claim 7, wherein the TNF receptor superfamily molecule is OPG. 8. The method of claim 7, wherein at least a portion of the cysteine rich domain of wild-type DcR3 is selected from all or part of CRD1, all or part of CRD2, all or part of CRD3, and all or part of CRD4,
At least a portion of the cysteine-rich domain of the TNF receptor superfamily molecule is all or part of CRD1, all or part of CRD2, all or part of CRD3, and all or part of CRD4 DcR3 variant. 15. The method of claim 14, wherein the first chimeric cysteine rich region comprises:
a substitution of a portion of CRD1 of the wild-type DcR3 in a portion corresponding to a portion of CRD1 of the wild-type DcR3 in CRD1 of the TNF receptor superfamily molecule;
substitution of all of CRD1 of said wild-type DcR3 in all of CRD1 of said TNF receptor superfamily molecule;
a substitution of a portion of CRD2 of said wild-type DcR3 in a portion corresponding to a portion of CRD2 of said wild-type DcR3 in CRD2 of said TNF receptor superfamily molecule;
substitution of all of CRD2 of said wild-type DcR3 in all of CRD2 of said TNF receptor superfamily molecule;
a substitution of a portion of CRD3 of said wild-type DcR3 in a portion corresponding to a portion of CRD3 of said wild-type DcR3 in CRD3 of said TNF receptor superfamily molecule;
substitution of all of CRD3 of said wild-type DcR3 in all of CRD3 of said TNF receptor superfamily molecule;
a substitution of a portion of CRD4 of said wild-type DcR3 in a portion corresponding to a portion of CRD4 of said wild-type DcR3 in CRD4 of said TNF receptor superfamily molecule, and
Substitution of all of CRD4 of said wild-type DcR3 in all of CRD4 of said TNF receptor superfamily molecule
DcR3 variant having one or more substitutions selected from. The DcR3 variant according to claim 15, wherein in the first chimeric cysteine-rich region, part or all of CRD2 of the wild-type DcR3 is maintained. The DcR3 variant according to claim 15, wherein in the first chimeric cysteine-rich region, part or all of CRD3 of the wild-type DcR3 is maintained. 15. The method according to claim 14, wherein the first chimeric cysteine-rich region comprises the following amino acid sequences (a), (b), (c) or (d), and the second chimeric cysteine-rich region comprises: DcR3 variant comprising the amino acid sequence of (e) below.
(a) In the amino acid sequence of the cysteine-rich region of wild-type DcR3, CRD1 of wild-type DcR3 is substituted with CRD1 of OPG
(b) in the amino acid sequence of the cysteine-rich region of wild-type DcR3, CRD4 of wild-type DcR3 is substituted with CRD4 of OPG
(c) an amino acid sequence in which CRD1 of wild-type DcR3 is substituted with CRD1 of OPG in the amino acid sequence of the cysteine-rich region of wild-type DcR3, and CRD4 of wild-type DcR3 is substituted with CRD4 of OPG
(d) in the amino acid sequence of (a), (b) or (c), the 103rd to 123rd position from the N-terminus is substituted with a corresponding part in the amino acid sequence of the cysteine-rich domain of OPG
(e) in the amino acid sequence of (a), (b), (c) or (d), 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence The amino acid sequence according to claim 18, wherein the amino acid sequence of (a) is an amino acid sequence comprising amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 26 or 50,
The amino acid sequence of (b) is an amino acid sequence comprising the amino acid at positions 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 28 or 52,
The amino acid sequence of (c) is an amino acid sequence comprising the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 30 or 54,
The DcR3 variant, wherein the amino acid sequence of (d) is an amino acid sequence comprising the amino acids 1 to 164 from the N-terminus in the amino acid sequence set forth in SEQ ID NO: 32 or 56. The amino acid sequence of claim 18, wherein the amino acid sequence of (e) is Glu at position 57, Arg at position 58, and Glu at position 60 from the N-terminus of the amino acid sequence of (a), (b), (c) or (d). One or two or more amino acids selected from the group consisting of Arg of DcR3 variant having a substitution with another amino acid. 19. The method according to claim 18, wherein the amino acid sequence of (e) is, from the N-terminus of the amino acid sequence of (a), (b), (c) or (d), Glu at position 57 and Arg at position 58; DcR3 variant having a substitution with an amino acid. The amino acid sequence of claim 18, wherein the amino acid sequence of (e) is Lys, Leu, Arg, Val of Glu at position 57 from the N-terminus of the amino acid sequence of (a), (b), (c) or (d). , Ala, Phe, His, He or Met, Arg at position 58 is replaced with Asp, Glu or Thr, and Arg at position 60 is selected from the group consisting of Lys. DcR3 variant. The amino acid sequence of claim 18, wherein the amino acid sequence of (e) is Lys, Leu, Arg, Val of Glu at position 57 from the N-terminus of the amino acid sequence of (a), (b), (c) or (d). , Ala, Phe, His, Ile or Met, and a DcR3 variant having a substitution of Asp, Glu or Thr in Arg at the 58th position from the N-terminus. The DcR3 variant according to claim 18, wherein the amino acid sequence of (e) has a substitution selected from the following (f) to (i).
(f) Substitution of Asn at positions 131 and 144 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid
(g) Substitution of Asn at positions 131, 144, and 157 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid
(h) Substitution of Thr at position 133 and Ser at position 146 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid
(i) Substitution of Thr at position 133, Ser at position 146, and Thr at position 159 from the N-terminus of the amino acid sequence of (b), (c) or (d) with another amino acid The DcR3 variant according to claim 18, wherein the amino acid sequence of (e) has a substitution selected from the following (f') to (i').
(f') Substitution of Asn at positions 131 and 144 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ser
(g') Substitution of Asn at positions 131, 144, and 157 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ser
(h') Substitution of Thr at position 133 and Ser at position 146 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ala
(i') Substitution of Thr at position 133, Ser at position 146, and Thr at position 159 from the N-terminus of the amino acid sequence of (b), (c) or (d) with Ala The method according to claim 18, wherein the amino acid sequence of (e) is SEQ ID NO: 58, 60, 62, 64, 66, 68, 70, 180, 182, 184, 186, 188, 270, 272, 274, 276, 278 , 280, 282, 284 or 286 of the amino acid sequence described in the amino acid sequence 1 to 164 amino acids from the N-terminal DcR3 variant. The heparan sulfate binding region of the wild-type DcR3 according to claim 7, wherein the DcR3 variant is bound to the C-terminal side of the first or second chimeric cysteine-rich region and the first or second chimeric cysteine-rich region A DcR3 variant comprising a part or all of a DcR3 variant, or a DcR3 variant comprising the first or second chimeric cysteine-rich region and not including the heparan sulfate binding region of the wild-type DcR3. The method of claim 27, wherein the DcR3 variant,
(I) the amino acid sequence set forth in SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44 or 46, or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence, and
(II) SEQ ID NOs: 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 180, 182, 184, 186, 188, 270, 272, 274, 276, 278, 280, 282 , 284 or 286, or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added amino acid sequence
DcR3 variant comprising any one amino acid sequence selected from. The amino acid sequence according to claim 7, wherein the DcR3 variant is a human IgG1, IgG2 or IgG4 antibody-derived Fc region, or in the amino acid sequence of the Fc region, one or several amino acids are deleted, substituted, inserted or added. DcR3 variant comprising a variant Fc region comprising. The DcR3 variant according to claim 29, wherein the Fc region or the variant Fc region is linked to the C-terminal side of the first or second chimeric cysteine-rich region via another region or a linker. The DcR3 variant according to claim 29, wherein the mutant Fc region has a Ser-substituted for Cys at position 220 indicated by the EU index in the amino acid sequence of the heavy chain of human IgG1. 32. The mutant Fc region according to claim 31, wherein, in the amino acid sequence of the heavy chain of human IgG1, Leu at position 234 indicated by the EU index is substituted with Ala, Leu at position 235 is substituted with Ala, and Gly at position 237 DcR3 variant having a substitution of Ala. The DcR3 variant according to claim 31, wherein the variant Fc region has a substitution of Ala for Asn at position 434 indicated by the EU index in the amino acid sequence of a heavy chain of human IgG1. The mutant Fc region according to claim 31, wherein, in the amino acid sequence of the heavy chain of human IgG1, Met at position 252 represented by the EU index is substituted with Tyr, Ser at position 254 is substituted with Thr, and Thr at position 256 DcR3 variant having a substitution with Glu. 30. The mutant Fc region according to claim 29, wherein, in the amino acid sequence of the heavy chain of human IgG4, Ser at position 228 represented by the EU index is substituted with Pro, Leu at position 235 is substituted with Glu, and Arg at position 409. DcR3 variant having a substitution of Lys. 30. The method of claim 29, wherein the DcR3 variant, SEQ ID NO: 72, 74, 156, 158, 160, 162, 164, 166, 311, 312 or 313 DcR3 dog comprising a variant Fc region comprising the amino acid sequence set forth variant. 30. The method of claim 29, wherein the DcR3 variant is SEQ ID NO: 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 150, 168, 170, 172, 174, 176, 178 , 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238 , 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 288, 290, 292, 294, 296, 298, 300, 302, 304, 314 , 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336 or 337. , or in the amino acid sequence, 1 to 30 amino acids are deleted, substituted, inserted or added DcR3 variant comprising an amino acid sequence. A DcR3 variant composition comprising the variant DcR3 according to claim 1 or 7. The composition according to claim 38, comprising a DcR3 variant having one or more N-glycosidic conjugated sugar chains and a DcR3 variant having no N-glycosidic conjugated sugar chain. A DNA encoding the DcR3 variant according to claim 1 or 7. A genetic recombinant vector containing the DNA according to claim 40. A transformant obtained by introducing the genetic recombinant vector according to claim 41 into a host cell. 43. The transformant according to claim 42, wherein the host cell is a cell derived from a mammal. 44. The transformant according to claim 43, wherein the mammalian-derived cell is a CHO cell. A DcR3 variant characterized in that the transformant according to claim 42 is cultured in a medium, the DcR3 variant according to claim 1 or 7 is produced and accumulated, and the DcR3 variant is purified from the obtained culture solution, or Method for preparing a DcR3 variant composition. A DcR3 variant or a DcR3 variant composition prepared using the production method according to claim 45. A pharmaceutical composition comprising the modified DcR3 or the modified DcR3 composition according to claim 1, 7 or 46 as an active ingredient. The pharmaceutical composition according to claim 47, which is a prophylactic or therapeutic agent for an autoimmune disease, an inflammatory disease, or an allergic disease. A method for preventing or treating an autoimmune disease, an inflammatory disease, or an allergic disease, comprising administering the pharmaceutical composition according to claim 47 to a patient in need of the prevention or treatment of an autoimmune disease, an inflammatory disease, or an allergic disease.

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